3,4-disubstituted 1h-pyrazole compounds and their use as cyclin dependent kinase and glycogen synthase kinase-3 modulators

ABSTRACT

The invention provides compounds of the formula (0) or salts or tautomers or N-oxides or solvates thereof, and combinations thereof with other anti-cancer agents, for use in the prophylaxis or treatment of disease states and conditions such as cancers mediated by cyclin-dependent kinase and glycogen synthase kinase-3.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.12/877,499, filed Sep. 8, 2010, which is a continuation of U.S.application Ser. No. 11/875,482, filed Oct. 19, 2007 (which issued asU.S. Pat. No. 7,825,140 on Nov. 2, 2010), which is a continuation ofU.S. Application No. 11,336,599, filed Jan. 20, 2006 (which issued asU.S. Pat. No. 7,385,059 on Jun. 10, 2008), which is a continuation ofPCT International Application PCT/GB2004/003179, filed Jul. 22, 2004,and published under PCT Article 21 (2) in English as WO 2005/012256 onFeb. 10, 2005. PCT/GB2004/003179 claimed benefit from U.S. ProvisionalApplications 60/489,046, filed Jul. 22, 2003, and 60/569,763, filed May10, 2004. PCT/GB2004/003179 also claimed priority from Britishapplication 0317127.9, filed Jul. 22, 2003. The entire contents of eachof the prior applications are incorporated herein by reference.

FIELD OF THE INVENTION

This invention relates to pyrazole compounds that inhibit or modulatethe activity of cyclin dependent kinases (CDK) and glycogen synthasekinase-3 (GSK-3), to the use of the compounds in the treatment orprophylaxis of disease states or conditions mediated by cyclin dependentkinases and glycogen synthase kinase-3, and to novel compounds havingcyclin dependent kinase or glycogen synthase kinase-3 inhibitory ormodulating activity. Also provided are pharmaceutical compositionscontaining the compounds and novel chemical intermediates.

BACKGROUND OF THE INVENTION

Protein kinases constitute a large family of structurally relatedenzymes that are responsible for the control of a wide variety of signaltransduction processes within the cell (Hardie, G. and Hanks, S. (1995)The Protein Kinase Facts Book. I and II, Academic Press, San Diego,Calif.). The kinases may be categorized into families by the substratesthey phosphorylate (e.g., protein-tyrosine, protein-serine/threonine,lipids, etc.). Sequence motifs have been identified that generallycorrespond to each of these kinase families (e.g., Hanks, S. K., Hunter,T., FASEB J., 9:576-596 (1995); Knighton, et al., Science, 253:407-414(1991); Hiles, et al., Cell, 70:419-429 (1992); Kunz, et al., Cell,73:585-596 (1993); Garcia-Bustos, et al., EMBO J., 13:2352-2361 (1994)).

Protein kinases may be characterized by their regulation mechanisms.These mechanisms include, for example, autophosphorylation,transphosphorylation by other kinases, protein-protein interactions,protein-lipid interactions, and protein-polynucleotide interactions. Anindividual protein kinase may be regulated by more than one mechanism.

Kinases regulate many different cell processes including, but notlimited to, proliferation, differentiation, apoptosis, motility,transcription, translation and other signalling processes, by addingphosphate groups to target proteins. These phosphorylation events act asmolecular on/off switches that can modulate or regulate the targetprotein biological function. Phosphorylation of target proteins occursin response to a variety of extracellular signals (hormones,neurotransmitters, growth and differentiation factors, etc.), cell cycleevents, environmental or nutritional stresses, etc. The appropriateprotein kinase functions in signalling pathways to activate orinactivate (either directly or indirectly), for example, a metabolicenzyme, regulatory protein, receptor, cytoskeletal protein, ion channelor pump, or transcription factor. Uncontrolled signalling due todefective control of protein phosphorylation has been implicated in anumber of diseases, including, for example, inflammation, cancer,allergy/asthma, disease and conditions of the immune system, disease andconditions of the central nervous system, and angiogenesis.

The process of eukaryotic cell division may be broadly divided into aseries of sequential phases termed G1, S, G2 and M. Correct progressionthrough the various phases of the cell cycle has been shown to becritically dependent upon the spatial and temporal regulation of afamily of proteins known as cyclin dependent kinases (CDKs) and adiverse set of their cognate protein partners termed cyclins. CDKs arecdc2 (also known as CDK1) homologous serine-threonine kinase proteinsthat are able to utilise ATP as a substrate in the phosphorylation ofdiverse polypeptides in a sequence dependent context. Cyclins are afamily of proteins characterised by a homology region, containingapproximately 100 amino acids, termed the “cyclin box” which is used inbinding to, and defining selectivity for, specific CDK partner proteins.

Modulation of the expression levels, degradation rates, and activationlevels of various CDKs and cyclins throughout the cell cycle leads tothe cyclical formation of a series of CDK/cyclin complexes, in which theCDKs are enzymatically active. The formation of these complexes controlspassage through discrete cell cycle checkpoints and thereby enables theprocess of cell division to continue. Failure to satisfy thepre-requisite biochemical criteria at a given cell cycle checkpoint,i.e. failure to form a required CDK/cyclin complex, can lead to cellcycle arrest and/or cellular apoptosis. Aberrant cellular proliferation,as manifested in cancer, can often be attributed to loss of correct cellcycle control. Inhibition of CDK enzymatic activity therefore provides ameans by which abnormally dividing cells can have their divisionarrested and/or be killed. The diversity of CDKs, and CDK complexes, andtheir critical roles in mediating the cell cycle, provides a broadspectrum of potential therapeutic targets selected on the basis of adefined biochemical rationale.

Progression from the G1 phase to the S phase of the cell cycle isprimarily regulated by CDK2, CDK3, CDK4 and CDK6 via association withmembers of the D and E type cyclins. The D-type cyclins appearinstrumental in enabling passage beyond the G1 restriction point, whereas the CDK2/cyclin E complex is key to the transition from the G1 to Sphase. Subsequent progression through S phase and entry into G2 isthought to require the CDK2/cyclin A complex. Both mitosis, and the G2to M phase transition which triggers it, are regulated by complexes ofCDK1 and the A and B type cyclins.

During G1 phase Retinoblastoma protein (Rb), and related pocket proteinssuch as p130, are substrates for CDK(2, 4, & 6)/cyclin complexes.Progression through G1 is in part facilitated by hyperphosphorylation,and thus inactivation, of Rb and p130 by the CDK(4/6)/cyclin-Dcomplexes. Hyperphosphorylation of Rb and p130 causes the release oftranscription factors, such as E2F, and thus the expression of genesnecessary for progression through G1 and for entry into S-phase, such asthe gene for cyclin E. Expression of cyclin E facilitates formation ofthe CDK2/cyclin E complex which amplifies, or maintains, E2F levels viafurther phosphorylation of Rb. The CDK2/cyclin E complex alsophosphorylates other proteins necessary for DNA replication, such asNPAT, which has been implicated in histone biosynthesis. G1 progressionand the G1/S transition are also regulated via the mitogen stimulatedMyc pathway, which feeds into the CDK2/cyclin E pathway. CDK2 is alsoconnected to the p53 mediated DNA damage response pathway via p53regulation of p21 levels. p21 is a protein inhibitor of CDK2/cyclin Eand is thus capable of blocking, or delaying, the G1/S transition. TheCDK2/cyclin E complex may thus represent a point at which biochemicalstimuli from the Rb, Myc and p53 pathways are to some degree integrated.CDK2 and/or the CDK2/cyclin E complex therefore represent good targetsfor therapeutics designed at arresting, or recovering control of, thecell cycle in aberrantly dividing cells.

The exact role of CDK3 in the cell cycle is not clear. As yet no cognatecyclin partner has been identified, but a dominant negative form of CDK3delayed cells in G1, thereby suggesting that CDK3 has a role inregulating the G1/S transition.

Although most CDKs have been implicated in regulation of the cell cyclethere is evidence that certain members of the CDK family are involved inother biochemical processes. This is exemplified by CDK5 which isnecessary for correct neuronal development and which has also beenimplicated in the phosphorylation of several neuronal proteins such asTau, NUDE-1, synapsin1, DARPP32 and the Munc18/Syntaxin1 A complex.Neuronal CDK5 is conventionally activated by binding to the p35/p39proteins. CDK5 activity can, however, be deregulated by the binding ofp25, a truncated version of p35. Conversion of p35 to p25, andsubsequent deregulation of CDK5 activity, can be induced by ischemia,excitotoxicity, and β-amyloid peptide. Consequently p25 has beenimplicated in the pathogenesis of neurodegenerative diseases, such asAlzheimer's, and is therefore of interest as a target for therapeuticsdirected against these diseases.

CDK7 is a nuclear protein that has cdc2 CAK activity and binds to cyclinH. CDK7 has been identified as component of the TFIIH transcriptionalcomplex which has RNA polymerase II C-terminal domain (CTD) activity.This has been associated with the regulation of HIV-1 transcription viaa Tat-mediated biochemical pathway. CDK8 binds cyclin C and has beenimplicated in the phosphorylation of the CTD of RNA polymerase II.Similarly the CDK9/cyclin-T1 complex (P-TEFb complex) has beenimplicated in elongation control of RNA polymerase II. PTEF-b is alsorequired for activation of transcription of the HIV-1 genome by theviral transactivator Tat through its interaction with cyclin T1. CDK7,CDK8, CDK9 and the P-TEFb complex are therefore potential targets foranti-viral therapeutics.

At a molecular level mediation of CDK/cyclin complex activity requires aseries of stimulatory and inhibitory phosphorylation, ordephosphorylation, events. CDK phosphorylation is performed by a groupof CDK activating kinases (CAKs) and/or kinases such as wee1, Myt1 andMik1. Dephosphorylation is performed by phosphatases such as cdc25(a &c), pp2a, or KAP.

CDK/cyclin complex activity may be further regulated by two families ofendogenous cellular proteinaceous inhibitors: the Kip/Cip family, or theINK family. The INK proteins specifically bind CDK4 and CDK6. p16^(ink4)(also known as MTS1) is a potential tumour suppressor gene that ismutated, or deleted, in a large number of primary cancers. The Kip/Cipfamily contains proteins such as p21^(Cip1,Waf1), p27^(Kip1) andp57^(Kip2). As discussed previously p21 is induced by p53 and is able toinactivate the CDK2/cyclin(E/A) and CDK4/cyclin(D1/D2/D3) complexes.Atypically low levels of p27 expression have been observed in breast,colon and prostate cancers. Conversely over expression of cyclin E insolid tumours has been shown to correlate with poor patient prognosis.Over expression of cyclin D1 has been associated with oesophageal,breast, squamous, and non-small cell lung carcinomas.

The pivotal roles of CDKs, and their associated proteins, inco-ordinating and driving the cell cycle in proliferating cells havebeen outlined above. Some of the biochemical pathways in which CDKs playa key role have also been described. The development of monotherapiesfor the treatment of proliferative disorders, such as cancers, usingtherapeutics targeted generically at CDKs, or at specific CDKs, istherefore potentially highly desirable. CDK inhibitors could conceivablyalso be used to treat other conditions such as viral infections,autoimmune diseases and neuro-degenerative diseases, amongst others. CDKtargeted therapeutics may also provide clinical benefits in thetreatment of the previously described diseases when used in combinationtherapy with either existing, or new, therapeutic agents. CDK targetedanticancer therapies could potentially have advantages over many currentantitumour agents as they would not directly interact with DNA andshould therefore reduce the risk of secondary tumour development.

Glycogen Synthase Kinase-3 (GSK3) is a serine-threonine kinase thatoccurs as two ubiquitously expressed isoforms in humans (GSK3α & betaGSK3β). GSK3 has been implicated as having roles in embryonicdevelopment, protein synthesis, cell proliferation, celldifferentiation, microtubule dynamics, cell motility and cellularapoptosis. As such GSK3 has been implicated in the progression ofdisease states such as diabetes, cancer, Alzheimer's disease, stroke,epilepsy, motor neuron disease and/or head trauma. Phylogenetically GSK3is most closely related to the cyclin dependent kinases (CDKs).

The consensus peptide substrate sequence recognised by GSK3 is(Ser/Thr)-X—X—X-(pSer/pThr), where X is any amino acid (at positions(n+1), (n+2), (n+3)) and pSer and pThr are phospho-serine andphospho-threonine respectively (n+4). GSK3 phosphorylates the firstserine, or threonine, at position (n). Phospho-serine, orphospho-threonine, at the (n+4) position appear necessary for primingGSK3 to give maximal substrate turnover. Phosphorylation of GSK3α atSer21, or GSK3β at Ser9, leads to inhibition of GSK3. Mutagenesis andpeptide competition studies have led to the model that thephosphorylated N-terminus of GSK3 is able to compete withphospho-peptide substrate (S/TXXXpS/pT) via an autoinhibitory mechanism.There are also data suggesting that GSK3α and GSKβ may be subtlyregulated by phosphorylation of tyrosines 279 and 216 respectively.Mutation of these residues to a Phe caused a reduction in in vivo kinaseactivity. The X-ray crystallographic structure of GSK3β has helped toshed light on all aspects of GSK3 activation and regulation.

GSK3 forms part of the mammalian insulin response pathway and is able tophosphorylate, and thereby inactivate, glycogen synthase. Upregulationof glycogen synthase activity, and thereby glycogen synthesis, throughinhibition of GSK3, has thus been considered a potential means ofcombating type II, or non-insulin-dependent diabetes mellitus (NIDDM): acondition in which body tissues become resistant to insulin stimulation.The cellular insulin response in liver, adipose, or muscle tissues, istriggered by insulin binding to an extracellular insulin receptor. Thiscauses the phosphorylation, and subsequent recruitment to the plasmamembrane, of the insulin receptor substrate (IRS) proteins. Furtherphosphorylation of the IRS proteins initiates recruitment ofphosphoinositide-3 kinase (PI3K) to the plasma membrane where it is ableto liberate the second messenger phosphatidylinosityl3,4,5-trisphosphate (PIP3). This facilitates co-localisation of3-phosphoinositide-dedependent protein kinase 1 (PDK1) and proteinkinase B (PKB or Akt) to the membrane, where PDK1 activates PKB. PKB isable to phosphorylate, and thereby inhibit, GSK3α and/or GSKβ throughphosphorylation of Ser9, or ser21, respectively. The inhibition of GSK3then triggers upregulation of glycogen synthase activity. Therapeuticagents able to inhibit GSK3 may thus be able to induce cellularresponses akin to those seen on insulin stimulation. A further in vivosubstrate of GSK3 is the eukaryotic protein synthesis initiation factor2B (eIF2B). eIF2B is inactivated via phosphorylation and is thus able tosuppress protein biosynthesis. Inhibition of GSK3, e.g. by inactivationof the “mammalian target of rapamycin” protein (mTOR), can thusupregulate protein biosynthesis. Finally there is some evidence forregulation of GSK3 activity via the mitogen activated protein kinase(MAPK) pathway through phosphorylation of GSK3 by kinases such asmitogen activated protein kinase activated protein kinase 1 (MAPKAP-K1or RSK). These data suggest that GSK3 activity may be modulated bymitogenic, insulin and/or amino acid stimulii.

It has also been shown that GSK3β is a key component in the vertebrateWnt signalling pathway. This biochemical pathway has been shown to becritical for normal embryonic development and regulates cellproliferation in normal tissues. GSK3 becomes inhibited in response toWnt stimulii. This can lead to the de-phosphorylation of GSK3 substratessuch as Axin, the adenomatous polyposis coli (APC) gene product andβ-catenin. Aberrant regulation of the Wnt pathway has been associatedwith many cancers. Mutations in APC, and/or β-catenin, are common incolorectal cancer and other tumours. β-catenin has also been shown to beof importance in cell adhesion. Thus GSK3 may also modulate cellularadhesion processes to some degree. Apart from the biochemical pathwaysalready described there are also data implicating GSK3 in the regulationof cell division via phosphorylation of cyclin-D1, in thephosphorylation of transcription factors such as c-Jun, CCAAT/enhancerbinding protein α (C/EBPα), c-Myc and/or other substrates such asNuclear Factor of Activated T-cells (NFATc), Heat Shock Factor-1 (HSF-1)and the c-AMP response element binding protein (CREB). GSK3 also appearsto play a role, albeit tissue specific, in regulating cellularapoptosis. The role of GSK3 in modulating cellular apoptosis, via apro-apoptotic mechanism, may be of particular relevance to medicalconditions in which neuronal apoptosis can occur. Examples of these arehead trauma, stroke, epilepsy, Alzheimer's and motor neuron diseases,progressive supranuclear palsy, corticobasal degeneration, and Pick'sdisease. In vitro it has been shown that GSK3 is able tohyper-phosphorylate the microtubule associated protein Tau.Hyperphosphorylation of Tau disrupts its normal binding to microtubulesand may also lead to the formation of intra-cellular Tau filaments. Itis believed that the progressive accumulation of these filaments leadsto eventual neuronal dysfunction and degeneration. Inhibition of Tauphosphorylation, through inhibition of GSK3, may thus provide a means oflimiting and/or preventing neurodegenerative effects.

WO 02/34721 from Du Pont discloses a class of indeno[1,2-c]pyrazol-4-ones as inhibitors of cyclin dependent kinases.

WO 01/81348 from Bristol Myers Squibb describes the use of 5-thio-,sulphinyl- and sulphonylpyrazolo[3,4-b]-pyridines as cyclin dependentkinase inhibitors.

WO 00/62778 also from Bristol Myers Squibb discloses a class of proteintyrosine kinase inhibitors.

WO 01/72745A1 from Cyclacel describes 2-substituted4-heteroaryl-pyrimidines and their preparation, pharmaceuticalcompositions containing them and their use as inhibitors ofcyclin-dependant kinases (CDKs) and hence their use in the treatment ofproliferative disorders such as cancer, leukaemia, psoriasis and thelike.

WO 99/21845 from Agouron describes 4-aminothiazole derivatives forinhibiting cyclin-dependent kinases (CDKs), such as CDK1, CDK2, CDK4,and CDK6. The invention is also directed to the therapeutic orprophylactic use of pharmaceutical compositions containing suchcompounds and to methods of treating malignancies and other disorders byadministering effective amounts of such compounds.

WO 01/53274 from Agouron discloses as CDK kinase inhibitors a class ofcompounds which can comprise an amide-substituted benzene ring linked toan N-containing heterocyclic group.

WO 01/98290 (Pharmacia & Upjohn) discloses a class of3-aminocarbonyl-2-carboxamido thiophene derivatives as protein kinaseinhibitors.

WO 01/53268 and WO 01/02369 from Agouron disclose compounds that mediateor inhibit cell proliferation through the inhibition of protein kinasessuch as cyclin dependent kinase or tyrosine kinase. The Agouroncompounds have an aryl or heteroaryl ring attached directly or though aCH═CH or CH═N group to the 3-position of an indazole ring.

WO 00/39108 and WO 02/00651 (both to Du Pont Pharmaceuticals) describeheterocyclic compounds that are inhibitors of trypsin-like serineprotease enzymes, especially factor Xa and thrombin. The compounds arestated to be useful as anticoagulants or for the prevention ofthromboembolic disorders.

US 2002/0091116 (Zhu et al.), WO 01/19798 and WO 01/64642 each disclosediverse groups of heterocyclic compounds as inhibitors of Factor Xa.Some 1-substituted pyrazole carboxamides are disclosed and exemplified.

U.S. Pat. No. 6,127,382, WO 01/70668, WO 00/68191, WO 97/48672, WO97/19052 and WO 97/19062 (all to Allergan) each describe compoundshaving retinoid-like activity for use in the treatment of varioushyperproliferative diseases including cancers.

WO 02/070510 (Bayer) describes a class of amino-dicarboxylic acidcompounds for use in the treatment of cardiovascular diseases. Althoughpyrazoles are mentioned generically, there are no specific examples ofpyrazoles in this document.

WO 97/03071 (Knoll AG) discloses a class of heterocyclyl-carboxamidederivatives for use in the treatment of central nervous systemdisorders. Pyrazoles are mentioned generally as examples of heterocyclicgroups but no specific pyrazole compounds are disclosed or exemplified.

WO 97/40017 (Novo Nordisk) describes compounds that are modulators ofprotein tyrosine phosphatases.

WO 03/020217 (Univ. Connecticut) discloses a class of pyrazole3-carboxamides as cannabinoid receptor modulators for treatingneurological conditions. It is stated (page 15) that the compounds canbe used in cancer chemotherapy but it is not made clear whether thecompounds are active as anti-cancer agents or whether they areadministered for other purposes.

WO 01/58869 (Bristol Myers Squibb) discloses cannabinoid receptormodulators that can be used inter alia to treat a variety of diseases.The main use envisaged is the treatment of respiratory diseases,although reference is made to the treatment of cancer.

WO 01/02385 (Aventis Crop Science) discloses1-(quinoline-4-yl)-1H-pyrazole derivatives as fungicides.1-Unsubstituted pyrazoles are disclosed as synthetic intermediates.

WO 2004/039795 (Fujisawa) discloses amides containing a 1-substitutedpyrazole group as inhibitors of apolipoprotein B secretion. Thecompounds are stated to be useful in treating such conditions ashyperlipidemia.

WO 2004/000318 (Cellular Genomics) discloses various amino-substitutedmonocycles as kinase modulators. None of the exemplified compounds arepyrazoles.

SUMMARY OF THE INVENTION

The invention provides compounds that have cyclin dependent kinaseinhibiting or modulating activity, and which it is envisaged will beuseful in preventing or treating disease states or conditions mediatedby the kinases.

Thus, for example, it is envisaged that the compounds of the inventionwill be useful in alleviating or reducing the incidence of cancer.

Accordingly, in one aspect, the invention provides the use of a compoundfor the manufacture of a medicament for the prophylaxis or treatment ofa disease state or condition mediated by a cyclin dependent kinase, thecompound having the formula (0):

or salts or tautomers or N-oxides or solvates thereof;wherein

-   -   X is a group R¹-A-NR⁴— or a 5- or 6-membered carbocyclic or        heterocyclic ring;    -   A is a bond, SO₂, C═O, NR^(g)(C═O) or O(C═O) wherein R^(g) is        hydrogen or C₁₋₄ hydrocarbyl optionally substituted by hydroxy        or C₁₋₄ alkoxy;    -   Y is a bond or an alkylene chain of 1, 2 or 3 carbon atoms in        length;    -   R¹ is hydrogen; a carbocyclic or heterocyclic group having from        3 to 12 ring members; or a C₁₋₈ hydrocarbyl group optionally        substituted by one or more substituents selected from halogen        (e.g. fluorine), hydroxy, C₁₋₄ hydrocarbyloxy, amino, mono- or        di-C₁₋₄ hydrocarbylamino, and carbocyclic or heterocyclic groups        having from 3 to 12 ring members, and wherein 1 or 2 of the        carbon atoms of the hydrocarbyl group may optionally be replaced        by an atom or group selected from O, S, NH, SO, SO₂;    -   R² is hydrogen; halogen; C₁₋₄ alkoxy (e.g. methoxy); or a C₁₋₄        hydrocarbyl group optionally substituted by halogen (e.g.        fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy);    -   R³ is selected from hydrogen and carbocyclic and heterocyclic        groups having from 3 to 12 ring members; and    -   R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally        substituted by halogen (e.g. fluorine), hydroxyl or C₁₋₄ alkoxy        (e.g. methoxy).

In one embodiment, the invention provides the use of a compound for themanufacture of a medicament for the prophylaxis or treatment of adisease state or condition mediated by a cyclin dependent kinase, thecompound having the formula) (I⁰):

or salts or tautomers or N-oxides or solvates thereof;wherein

-   -   X is a group R¹-A-NR⁴— or a 5- or 6-membered carbocyclic or        heterocyclic ring;    -   A is a bond, C═O, NR^(g)(C═O) or O(C═O) wherein R^(g) is        hydrogen or C₁₋₄ hydrocarbyl optionally substituted by hydroxy        or C₁₋₄ alkoxy;    -   Y is a bond or an alkylene chain of 1, 2 or 3 carbon atoms in        length;    -   R¹ is hydrogen; a carbocyclic or heterocyclic group having from        3 to 12 ring members; or a C₁₋₈ hydrocarbyl group optionally        substituted by one or more substituents selected from halogen        (e.g. fluorine), hydroxy, C₁₋₄ hydrocarbyloxy, amino, mono- or        di-C₁₋₄ hydrocarbylamino, and carbocyclic or heterocyclic groups        having from 3 to 12 ring members, and wherein 1 or 2 of the        carbon atoms of the hydrocarbyl group may optionally be replaced        by an atom or group selected from O, S, NH, SO, SO₂;    -   R² is hydrogen; halogen; C₁₋₄ alkoxy (e.g. methoxy); or a C₁₋₄        hydrocarbyl group optionally substituted by halogen (e.g.        fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy);    -   R³ is selected from hydrogen and carbocyclic and heterocyclic        groups having from 3 to 12 ring members; and    -   R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally        substituted by halogen (e.g. fluorine), hydroxyl or C₁₋₄ alkoxy        (e.g. methoxy).

The invention also provides the use of a compound for the manufacture ofa medicament for the prophylaxis or treatment of a disease state orcondition mediated by a cyclin dependent kinase, the compound having theformula (I):

or salts or tautomers or N-oxides or solvates thereof;wherein

-   -   X is a group R¹-A-NR⁴—;    -   A is a bond, C═O, NR^(g)(C═O) or O(C═O) wherein R^(g) is        hydrogen or C₁₋₄ hydrocarbyl optionally substituted by hydroxy        or C₁₋₄ alkoxy;    -   Y is a bond or an alkylene chain of 1, 2 or 3 carbon atoms in        length;    -   R¹ is hydrogen; a carbocyclic or heterocyclic group having from        3 to 12 ring members; or a C₁₋₈ hydrocarbyl group optionally        substituted by one or more substituents selected from halogen        (e.g. fluorine), hydroxy, C₁₋₄ hydrocarbyloxy, amino, mono- or        di-C₁₋₄ hydrocarbylamino, and carbocyclic or heterocyclic groups        having from 3 to 12 ring members, and wherein 1 or 2 of the        carbon atoms of the hydrocarbyl group may optionally be replaced        by an atom or group selected from O, S, NH, SO, SO₂;    -   R² is hydrogen; halogen; C₁₋₄ alkoxy (e.g. methoxy); or a C₁₋₄        hydrocarbyl group optionally substituted by halogen (e.g.        fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy);    -   R³ is selected from hydrogen and carbocyclic and heterocyclic        groups having from 3 to 12 ring members; and    -   R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally        substituted by halogen (e.g. fluorine), hydroxyl or C₁₋₄ alkoxy        (e.g. methoxy).

Any one or more of the following optional provisos, in any combination,may apply to the compounds of formulae (0), (I⁰), (I) and sub-groupsthereof:

(a-i) When A is a bond and Y—R³ is an alkyl, cycloalkyl, optionallysubstituted phenyl or optionally substituted phenylalkyl, then R¹ isother than a substituted or unsubstituted dihydronaphthalene,dihydrochroman, dihydrothiochroman, tetrahydroquinoline ortetrahydrobenzfuranyl group.(a-ii) X and R³ are each other than a moiety containing a maleimidegroup wherein the maleimide group has nitrogen atoms attached to the 3-and 4-positions thereof.(a-iii) R¹ is other than a moiety containing a purine nucleoside group.(a-iv) X and R³ are each other than a moiety containing acyclobutene-1,2-dione group wherein the cyclobutene-1,2-dione group hasnitrogen atoms attached to the 3- and 4-positions thereof.(a-v) R³ is other than a moiety containing a 4-monosubstituted or4,5-disubstituted 2-pyridyl or 2-pyrimidinyl group or a5-monosubstituted or 5,6-disubstituted 1,2,4-triazin-3-yl or3-pyridazinyl group.(a-vi) X and R³ are each other than a moiety containing a substituted orunsubstituted pyrazol-3-ylamine group linked to a substituted orunsubstituted pyridine, diazine or triazine group.(a-vii) When A is C═O and Y—R³ is an alkyl, cycloalkyl, optionallysubstituted phenyl or optionally substituted phenylalkyl group, then R¹is other than a substituted or unsubstituted tetrahydronaphthalene,tetrahydroquinolinyl, tetrahydrochromanyl or tetrahydrothiochromanylgroup.(a-viii) When R³ is H and A is a bond, R¹ is other than a moietycontaining a bis-aryl, bis-heteroaryl or aryl heteroaryl group.(a-ix) R³ is other than a moiety containing a1,2,8,8a-tetrahydro-7-methyl-cyclopropa[c]pyrrolo[3,2,e]indole-4-(5H)-onegroup.(a-x) When Y is a bond, R³ is hydrogen, A is CO and R¹ is a substitutedphenyl group, each substituent on the phenyl group is other than a groupCH₂—P(O)R^(x)R^(y) where R^(x) and R^(y) are each selected from alkoxyand phenyl groups.(a-xi) X is other than4-(tert-butyloxycarbonylamino)-3-methylimidazol-2-ylcarbonylamino.

In another aspect, the invention provides, for use in medicine, asub-group of compounds of the formula (I) represented by the generalformula (Ia):

or salts or tautomers or N-oxides or solvates thereof;wherein

-   -   X is a group R¹-A-NR⁴—;    -   A is a bond, C═O, NR^(g)(C═O) or O(C═O) wherein R^(g) is        hydrogen or C₁₋₄ hydrocarbyl optionally substituted by hydroxy        or C₁₋₄ alkoxy;    -   Y is a bond or an alkylene chain of 1, 2 or 3 carbon atoms in        length;    -   R¹ is a carbocyclic or heterocyclic group having from 3 to 12        ring members; or a C₁₋₈ hydrocarbyl group optionally substituted        by one or more substituents selected from fluorine, hydroxy,        C₁₋₄ hydrocarbyloxy, amino, mono- or di-C₁₋₄ hydrocarbylamino,        and carbocyclic or heterocyclic groups having from 3 to 12 ring        members, and wherein 1 or 2 of the carbon atoms of the        hydrocarbyl group may optionally be replaced by an atom or group        selected from O, S, NH, SO, SO₂;    -   R² is hydrogen; halogen; C₁₋₄ alkoxy (e.g. methoxy); or a C₁₋₄        hydrocarbyl group optionally substituted by halogen (e.g.        fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy);    -   R³ is selected from hydrogen and carbocyclic and heterocyclic        groups having from 3 to 12 ring members; and    -   R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally        substituted by halogen (e.g. fluorine), hydroxyl or C₁₋₄ alkoxy        (e.g. methoxy).

Any one or more of the following optional provisos, in any combination,may apply to the compounds of formula (Ia) and sub-groups thereof:

Provisos (a-i) to (a-xi) above.

(b-i) R³ is other than a bridged azabicyclo group.(b-ii) When A is a bond, then R³ is other than a moiety containing anunsubstituted or substituted phenyl group having attached to an orthoposition thereof, a substituted or unsubstituted carbamoyl orthiocarbamoyl group.(b-iii) When A is a bond, then R³ is other than a moiety containing anisoquinoline or quinoxaline group each having attached thereto asubstituted or unsubstituted piperidine or piperazine ring.(b-iv) When A is a bond and R¹ is an alkyl group, then R³ is other thana moiety containing a thiatriazine group.(b-v) When R¹ or R³ contain a moiety in which a heterocyclic ring havingan S(═O)₂ ring member is fused to a carbocyclic ring, the saidcarbocyclic ring is other than a substituted or unsubstituted benzenering(b-vi) When A is a bond, R¹ is other than an arylalkyl, heteroarylalkylor piperidinylalkyl group each having attached thereto a substituentselected from cyano, and substituted or unsubstituted amino, aminoalkyl,amidine, guanidine, and carbamoyl groups.(b-vii) When X is a group R¹-A-NR⁴—, A is a bond and R¹ is anon-aromatic group, then R³ is other than a six membered monocyclic arylor heteroaryl group linked directly to a 5,6-fused bicyclic heteroarylgroup.

In a further aspect, the invention provides a sub-group of novelcompounds of the formulae (I) and (Ia) as defined herein, the novelcompounds being represented by the formula (Ib):

or salts or tautomers or N-oxides or solvates thereof;wherein

-   -   X is a group R¹-A-NR⁴—;    -   A is a bond, C═O, NR^(g)(C═O) or O(C═O) wherein R^(g) is        hydrogen or C₁₋₄ hydrocarbyl optionally substituted by hydroxy        or C₁₋₄ alkoxy;    -   Y is a bond or an alkylene chain of 1, 2 or 3 carbon atoms in        length;    -   R¹ is a carbocyclic or heterocyclic group having from 3 to 12        ring members; or a C₁₋₈ hydrocarbyl group optionally substituted        by one or more substituents selected from fluorine, hydroxy,        C₁₋₄ hydrocarbyloxy, amino, mono- or di-C₁₋₄ hydrocarbylamino,        and carbocyclic or heterocyclic groups having from 3 to 12 ring        members, and wherein 1 or 2 of the carbon atoms of the        hydrocarbyl group may optionally be replaced by an atom or group        selected from O, S, NH, SO, SO₂;    -   R² is hydrogen; halogen; C₁₋₄ alkoxy (e.g. methoxy); or a C₁₋₄        hydrocarbyl group optionally substituted by halogen (e.g.        fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy);    -   R³ is selected from carbocyclic and heterocyclic groups having        from 3 to 12 ring members; and    -   R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally        substituted by halogen (e.g. fluorine), hydroxyl or C₁₋₄        alkoxy(e.g. methoxy).

Any one or more of the following optional provisos, in any combination,may apply to the compounds of formula (Ib) and sub-groups thereof:

Provisos (a-i) to (a-vii), (a-ix) and (a-xi).

Provisos (b-i) to (b-vii).

(c-i) When A is a bond, R¹ is other than a substituted arylalkyl,heteroarylalkyl or piperidinylalkyl group.(c-ii) When X is an amino or alkylamino group and Y is a bond, R³ isother than a disubstituted thiazolyl group wherein one of thesubstituents is selected from cyano and fluoroalkyl.

The reference in proviso (a-iii) to a purine nucleoside group refers tosubstituted and unsubstituted purine groups having attached thereto amonosaccharide group (e.g. a pentose or hexose) or a derivative of amonosaccharide group, for example a deoxy monosaccharide group or asubstituted monosaccharide group.

The reference in proviso (b-i) to a bridged azabicyclo group refers tobicycloalkane bridged ring systems in which one of the carbon atoms ofthe bicycloalkane has been replaced by a nitrogen atom. In bridged ringsystems, two rings share more than two atoms, see for example AdvancedOrganic Chemistry, by Jerry March, 4^(th) Edition, Wiley Interscience,pages 131-133, 1992.

The invention also provides the use of a compound of the formulae (Ia)or (Ib) as defined herein for the manufacture of a medicament for theprophylaxis or treatment of a disease state or condition mediated by acyclin dependent kinase.

The provisos (a-i) to (a-x), (b-i) to (b-vii), (c-i) and (c-ii) informulae (I), (Ia) and (Ib) above refer to the disclosures in thefollowing prior art documents.

-   (a-i) US 2003/0166932, U.S. Pat. No. 6,127,382, U.S. Pat. No.    6,093,838-   (a-ii) WO 03/031440-   (a-iii) WO 03/014137-   (a-iv) WO 02/083624-   (a-v) WO 02/064586-   (a-vi) WO 02/22608, WO 02/22605, WO 02/22603 & WO 02/22601-   (a-vii) WO 97/48672, WO 97/19052-   (a-viii) WO 00/06169-   (a-ix) U.S. Pat. No. 5,502,068-   (a-x) JP 07188269-   (b-i) WO 03/040147-   (b-ii) WO 01/70671-   (b-iii) WO 01/32626-   (b-iv) WO 98/08845-   (b-v) WO 00/59902-   (b-vi) U.S. Pat. No. 6,020,357, WO 99/32454 & WO 98/28269-   (b-vii) WO 2004/012736-   (c-i) U.S. Pat. No. 6,020,357, WO 99/32454 & WO 98/28269-   (c-ii) US 2004/0082629

Any one or more of the foregoing optional provisos, (a-i) to (a-xi),(b-i) to (b-vii), (c-i) and (c-ii) in any combination, may also apply tothe compounds of formulae (Ib), (II), (III), (IV), (IVa), (Va), (Vb),(VIa), (VIb), (VII) or (VIII) and sub-groups thereof as defined herein.

The invention also provides:

-   -   The use of a compound of the formula (Ia), (Ib), (II), (III),        (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and        sub-groups thereof as defined herein for manufacture of a        medicament for the prophylaxis or treatment of a disease state        or condition mediated by a cyclin dependent kinase.    -   A method for alleviating or reducing the incidence of a disease        or condition comprising or arising from abnormal cell growth in        a mammal, which method comprises administering to the mammal a        compound of the formula (0), (I⁰), (I), (Ia), (Ib), (II), (III),        (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and        sub-groups thereof as defined herein in an amount effective in        inhibiting abnormal cell growth.    -   A method for alleviating or reducing the incidence of a disease        state or condition mediated by a cyclin dependent kinase or        glycogen synthase kinase-3, which method comprises administering        to a subject in need thereof a compound of the formula (0),        (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb),        (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as defined        herein.    -   A method for the prophylaxis or treatment of a disease state or        condition mediated by a cyclin dependent kinase, which method        comprises administering to a subject in need thereof a compound        of the formula (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV),        (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups        thereof as defined herein.    -   A method for treating a disease or condition comprising or        arising from abnormal cell growth in a mammal, which method        comprises administering to the mammal a compound of the formula        (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va),        (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as        defined herein in an amount effective in inhibiting abnormal        cell growth.    -   A method for treating a disease or condition comprising or        arising from abnormal cell growth in a mammal, the method        comprising administering to the mammal a compound of the formula        (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va),        (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as        defined herein in an amount effective to inhibit a cyclin        dependent kinase (e.g. CDK2).    -   A method of inhibiting a cyclin dependent kinase, which method        comprises contacting the kinase with a kinase-inhibiting        compound of the formula (0), (I⁰), (I), (Ia) (Ib), (II), (III),        (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and        sub-groups thereof as defined herein.    -   A method of modulating a cellular process (for example cell        division) by inhibiting the activity of a cyclin dependent        kinase using a compound of the formula (0), (I⁰), (I), (Ia),        (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII)        or (VIII) and sub-groups thereof as defined herein.

The compounds of the invention are also considered to be inhibitors ofglycogen synthase kinase-3 (GSK3) and, accordingly, the invention alsoprovides methods and uses of kinase inhibitors or modulators of theformula (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va),(Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as definedherein but wherein the kinase is glycogen synthase kinase-3.

In further aspects, the invention provides:

-   -   A pharmaceutical composition comprising a compound of the        formula (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa),        (VIb), (VII) or (VIII) and sub-groups thereof as defined herein        and a pharmaceutically acceptable carrier.    -   Compounds of the formula (Ib), (II), (III), (IV), (IVa), (Va),        (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as        defined herein for use in medicine.    -   The use of a compound of the formula (0), (I⁰), (I), (Ia), (Ib),        (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII)        or (VIII) and sub-groups thereof as defined herein, for the        manufacture of a medicament for the prophylaxis or treatment of        any one of the disease states or conditions disclosed herein.    -   A method for the treatment or prophylaxis of any one of the        disease states or consitions disclosed herein, which method        comprises administering to a patient (e.g. a patient in need        thereof) a compound (e.g. a therapeutically effective amount) of        the formula (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV),        (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups        thereof as defined herein.    -   A method for alleviating or reducing the incidence of a disease        state or condition disclosed herein, which method comprises        administering to a patient (e.g. a patient in need thereof) a        compound (e.g. a therapeutically effective amount) of the        formula (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa),        (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof        as defined herein.    -   A method for the diagnosis and treatment of a disease state or        condition mediated by a cyclin dependent kinase, which method        comprises (i) screening a patient to determine whether a disease        or condition from which the patient is or may be suffering is        one which would be susceptible to treatment with a compound        having activity against cyclin dependent kinases; and (ii) where        it is indicated that the disease or condition from which the        patient is thus susceptible, thereafter administering to the        patient a compound of the formula (0), (I⁰), (I), (Ia), (Ib),        (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII)        or (VIII) and sub-groups thereof as defined herein.    -   The use of a compound of the formula (0), (I⁰), (I), (Ia), (Ib),        (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII)        or (VIII) and sub-groups thereof as defined herein for the        manufacture of a medicament for the treatment or prophylaxis of        a disease state or condition in a patient who has been screened        and has been determined as suffering from, or being at risk of        suffering from, a disease or condition which would be        susceptible to treatment with a compound having activity against        cyclin dependent kinase.

In each of the foregoing uses, methods and other aspects of theinvention, as well as any aspects and embodiments of the invention asset out below, references to compounds of the formulae (0), (I⁰), (I),(Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or(VIII) and sub-groups thereof as defined herein include within theirscope the salts or solvates or tautomers or N-oxides of the compounds.

General Preferences and Definitions

The following general preferences and definitions shall apply to each ofthe moieties X, Y, R^(g), R¹ to R⁴ and any sub-definition, sub-group orembodiment thereof, unless the context indicates otherwise.

In this specification, references to formula (I) include formulae (0),(I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb),(VII) or (VIII) and sub-groups, examples or embodiments of formulae (0),(I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb),(VII) or (VIII) unless the context indicates otherwise.

Thus for example, references to inter alia therapeutic uses,pharmaceutical formulations and processes for making compounds, wherethey refer to formula (I), are also to be taken as referring to formulae(0), (I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa),(VIb), (VII) or (VIII) and sub-groups, examples or embodiments offormulae (0), (I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb),(VIa), (VIb), (VII) or (VIII).

Similarly, where preferences, embodiments and examples are given forcompounds of the formula (I), they are also applicable to formulae (0),(I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb),(VII) or (VIII) and sub-groups, examples or embodiments of formulae (0),(I⁰), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb),(VII) or (VIII) unless the context requires otherwise.

References to “carbocyclic” and “heterocyclic” groups as used hereinshall, unless the context indicates otherwise, include both aromatic andnon-aromatic ring systems. Thus, for example, the term “carbocyclic andheterocyclic groups” includes within its scope aromatic, non-aromatic,unsaturated, partially saturated and fully saturated carbocyclic andheterocyclic ring systems. In general, such groups may be monocyclic orbicyclic and may contain, for example, 3 to 12 ring members, moreusually 5 to 10 ring members. Examples of monocyclic groups are groupscontaining 3, 4, 5, 6, 7, and 8 ring members, more usually 3 to 7, andpreferably 5 or 6 ring members. Examples of bicyclic groups are thosecontaining 8, 9, 10, 11 and 12 ring members, and more usually 9 or 10ring members.

The carbocyclic or heterocyclic groups can be aryl or heteroaryl groupshaving from 5 to 12 ring members, more usually from 5 to 10 ringmembers. The term “aryl” as used herein refers to a carbocyclic grouphaving aromatic character and the term “heteroaryl” is used herein todenote a heterocyclic group having aromatic character. The terms “aryl”and “heteroaryl” embrace polycyclic (e.g. bicyclic) ring systems whereinone or more rings are non-aromatic, provided that at least one ring isaromatic. In such polycyclic systems, the group may be attached by thearomatic ring, or by a non-aromatic ring. The aryl or heteroaryl groupscan be monocyclic or bicyclic groups and can be unsubstituted orsubstituted with one or more substituents, for example one or moregroups R¹⁰ as defined herein.

The term “non-aromatic group” embraces unsaturated ring systems withoutaromatic character, partially saturated and fully saturated carbocyclicand heterocyclic ring systems. The terms “unsaturated” and “partiallysaturated” refer to rings wherein the ring structure(s) contains atomssharing more than one valence bond i.e. the ring contains at least onemultiple bond e.g. a C═C, C═C or N═C bond.

The term “fully saturated” refers to rings where there are no multiplebonds between ring atoms. Saturated carbocyclic groups includecycloalkyl groups as defined below. Partially saturated carbocyclicgroups include cycloalkenyl groups as defined below, for examplecyclopentenyl, cycloheptenyl and cyclooctenyl. A further example of acycloalkenyl group is cyclohexenyl.

Examples of heteroaryl groups are monocyclic and bicyclic groupscontaining from five to twelve ring members, and more usually from fiveto ten ring members. The heteroaryl group can be, for example, a fivemembered or six membered monocyclic ring or a bicyclic structure formedfrom fused five and six membered rings or two fused six membered ringsor, by way of a further example, two fused five membered rings. Eachring may contain up to about four heteroatoms typically selected fromnitrogen, sulphur and oxygen. Typically the heteroaryl ring will containup to 4 heteroatoms, more typically up to 3 heteroatoms, more usually upto 2, for example a single heteroatom. In one embodiment, the heteroarylring contains at least one ring nitrogen atom. The nitrogen atoms in theheteroaryl rings can be basic, as in the case of an imidazole orpyridine, or essentially non-basic as in the case of an indole orpyrrole nitrogen. In general the number of basic nitrogen atoms presentin the heteroaryl group, including any amino group substituents of thering, will be less than five.

Examples of five membered heteroaryl groups include but are not limitedto pyrrole, furan, thiophene, imidazole, furazan, oxazole, oxadiazole,oxatriazole, isoxazole, thiazole, isothiazole, pyrazole, triazole andtetrazole groups.

Examples of six membered heteroaryl groups include but are not limitedto pyridine, pyrazine, pyridazine, pyrimidine and triazine.

A bicyclic heteroaryl group may be, for example, a group selected from:

-   -   a) a benzene ring fused to a 5- or 6-membered ring containing 1,        2 or 3 ring heteroatoms;    -   b) a pyridine ring fused to a 5- or 6-membered ring containing        1, 2 or 3 ring heteroatoms;    -   c) a pyrimidine ring fused to a 5- or 6-membered ring containing        1 or 2 ring heteroatoms;    -   d) a pyrrole ring fused to a 5- or 6-membered ring containing 1,        2 or 3 ring heteroatoms;    -   e) a pyrazole ring fused to a 5- or 6-membered ring containing 1        or 2 ring heteroatoms;    -   f) an imidazole ring fused to a 5- or 6-membered ring containing        1 or 2 ring heteroatoms;    -   g) an oxazole ring fused to a 5- or 6-membered ring containing 1        or 2 ring heteroatoms;    -   h) an isoxazole ring fused to a 5- or 6-membered ring containing        1 or 2 ring heteroatoms;    -   i) a thiazole ring fused to a 5- or 6-membered ring containing 1        or 2 ring heteroatoms;    -   j) an isothiazole ring fused to a 5- or 6-membered ring        containing 1 or 2 ring heteroatoms;    -   k) a thiophene ring fused to a 5- or 6-membered ring containing        1, 2 or 3 ring heteroatoms;    -   l) a furan ring fused to a 5- or 6-membered ring containing 1, 2        or 3 ring heteroatoms;    -   m) an oxazole ring fused to a 5- or 6-membered ring containing 1        or 2 ring heteroatoms;    -   n) an isoxazole ring fused to a 5- or 6-membered ring containing        1 or 2 ring heteroatoms;    -   o) a cyclohexyl ring fused to a 5- or 6-membered ring containing        1, 2 or 3 ring heteroatoms; and    -   p) a cyclopentyl ring fused to a 5- or 6-membered ring        containing 1, 2 or 3 ring heteroatoms.

Particular examples of bicyclic heteroaryl groups containing a fivemembered ring fused to another five membered ring include but are notlimited to imidazothiazole (e.g. imidazo[2,1-b]thiazole) andimidazoimidazole (e.g. imidazo[1,2-a]imidazole).

Particular examples of bicyclic heteroaryl groups containing a sixmembered ring fused to a five membered ring include but are not limitedto benzfuran, benzthiophene, benzimidazole, benzoxazole, isobenzoxazole,benzisoxazole, benzthiazole, benzisothiazole, isobenzofuran, indole,isoindole, indolizine, indoline, isoindoline, purine (e.g., adenine,guanine), indazole, pyrazolopyrimidine (e.g. pyrazolo[1,5-a]pyrimidine),triazolopyrimidine (e.g. [1,2,4]triazolo[1,5-a]pyrimidine), benzodioxoleand pyrazolopyridine (e.g. pyrazolo[1,5-a]pyridine) groups.

Particular examples of bicyclic heteroaryl groups containing two fusedsix membered rings include but are not limited to quinoline,isoquinoline, chroman, thiochroman, chromene, isochromene, chroman,isochroman, benzodioxan, quinolizine, benzoxazine, benzodiazine,pyridopyridine, quinoxaline, quinazoline, cinnoline, phthalazine,naphthyridine and pteridine groups.

One sub-group of heteroaryl groups comprises pyridyl, pyrrolyl, furanyl,thienyl, imidazolyl, oxazolyl, oxadiazolyl, oxatriazolyl, isoxazolyl,thiazolyl, isothiazolyl, pyrazolyl, pyrazinyl, pyridazinyl, pyrimidinyl,triazinyl, triazolyl, tetrazolyl, quinolinyl, isoquinolinyl,benzfuranyl, benzthienyl, chromanyl, thiochromanyl, benzimidazolyl,benzoxazolyl, benzisoxazole, benzthiazolyl and benzisothiazole,isobenzofuranyl, indolyl, isoindolyl, indolizinyl, indolinyl,isoindolinyl, purinyl (e.g., adenine, guanine), indazolyl,benzodioxolyl, chromenyl, isochromenyl, isochromanyl, benzodioxanyl,quinolizinyl, benzoxazinyl, benzodiazinyl, pyridopyridinyl,quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, naphthyridinyl andpteridinyl groups.

Examples of polycyclic aryl and heteroaryl groups containing an aromaticring and a non-aromatic ring include tetrahydronaphthalene,tetrahydroisoquinoline, tetrahydroquinoline, dihydrobenzthiene,dihydrobenzfuran, 2,3-dihydro-benzo[1,4]dioxine, benzo[1,3]dioxole,4,5,6,7-tetrahydrobenzofuran, indoline and indane groups.

Examples of carbocyclic aryl groups include phenyl, naphthyl, indenyl,and tetrahydronaphthyl groups.

Examples of non-aromatic heterocyclic groups include unsubstituted orsubstituted (by one or more groups R¹⁰) heterocyclic groups having from3 to 12 ring members, typically 4 to 12 ring members, and more usuallyfrom 5 to 10 ring members. Such groups can be monocyclic or bicyclic,for example, and typically have from 1 to 5 heteroatom ring members(more usually 1,2, 3 or 4 heteroatom ring members) typically selectedfrom nitrogen, oxygen and sulphur.

When sulphur is present, it may, where the nature of the adjacent atomsand groups permits, exist as —S—, —S(O)— or —S(O)₂—.

The heterocylic groups can contain, for example, cyclic ether moieties(e.g. as in tetrahydrofuran and dioxane), cyclic thioether moieties(e.g. as in tetrahydrothiophene and dithiane), cyclic amine moieties(e.g. as in pyrrolidine), cyclic amide moieties (e.g. as inpyrrolidone), cyclic thioamides, cyclic thioesters, cyclic estermoieties (e.g. as in butyrolactone), cyclic sulphones (e.g. as insulpholane and sulpholene), cyclic sulphoxides, cyclic sulphonamides andcombinations thereof (e.g. morpholine and thiomorpholine and its S-oxideand S,S-dioxide). Further examples of heterocyclic groups are thosecontaining a cyclic urea moiety (e.g. as in imidazolidin-2-one),

In one sub-set of heterocyclic groups, the heterocyclic groups containcyclic ether moieties (e.g. as in tetrahydrofuran and dioxane), cyclicthioether moieties (e.g. as in tetrahydrothiophene and dithiane), cyclicamine moieties (e.g. as in pyrrolidine), cyclic sulphones (e.g. as insulpholane and sulpholene), cyclic sulphoxides, cyclic sulphonamides andcombinations thereof (e.g. thiomorpholine).

Examples of monocyclic non-aromatic heterocyclic groups include 5-, 6-and 7-membered monocyclic heterocyclic groups. Particular examplesinclude morpholine, piperidine (e.g. 1-piperidinyl, 2-piperidinyl,3-piperidinyl and 4-piperidinyl), pyrrolidine (e.g. 1-pyrrolidinyl,2-pyrrolidinyl and 3-pyrrolidinyl), pyrrolidone, pyran (2H-pyran or4H-pyran), dihydrothiophene, dihydropyran, dihydrofuran,dihydrothiazole, tetrahydrofuran, tetrahydrothiophene, dioxane,tetrahydropyran (e.g. 4-tetrahydro pyranyl), imidazoline,imidazolidinone, oxazoline, thiazoline, 2-pyrazoline, pyrazolidine,piperazine, and N-alkyl piperazines such as N-methyl piperazine. Furtherexamples include thiomorpholine and its S-oxide and S,S-dioxide(particularly thiomorpholine). Still further examples include azetidine,piperidone, piperazone, and N-alkyl piperidines such as N-methylpiperidine.

One preferred sub-set of non-aromatic heterocyclic groups consists ofsaturated groups such as azetidine, pyrrolidine, piperidine, morpholine,thiomorpholine, thiomorpholine S,S-dioxide, piperazine, N-alkylpiperazines, and N-alkyl piperidines.

Another sub-set of non-aromatic heterocyclic groups consists ofpyrrolidine, piperidine, morpholine, thiomorpholine, thiomorpholineS,S-dioxide, piperazine and N-alkyl piperazines such as N-methylpiperazine.

One particular sub-set of heterocyclic groups consists of pyrrolidine,piperidine, morpholine and N-alkyl piperazines (e.g. N-methylpiperazine), and optionally thiomorpholine.

Examples of non-aromatic carbocyclic groups include cycloalkane groupssuch as cyclohexyl and cyclopentyl, cycloalkenyl groups such ascyclopentenyl, cyclohexenyl, cycloheptenyl and cyclooctenyl, as well ascyclohexadienyl, cyclooctatetraene, tetrahydronaphthenyl and decalinyl.

Preferred non-aromatic carbocyclic groups are monocyclic rings and mostpreferably saturated monocyclic rings.

Typical examples are three, four, five and six membered saturatedcarbocyclic rings, e.g. optionally substituted cyclopentyl andcyclohexyl rings.

One sub-set of non-aromatic carboyclic groups includes unsubstituted orsubstituted (by one or more groups R¹⁰) monocyclic groups andparticularly saturated monocyclic groups, e.g. cycloalkyl groups.Examples of such cycloalkyl groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and cycloheptyl; more typically cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl, particularly cyclohexyl.

Further examples of non-aromatic cyclic groups include bridged ringsystems such as bicycloalkanes and azabicycloalkanes although suchbridged ring systems are generally less preferred. By “bridged ringsystems” is meant ring systems in which two rings share more than twoatoms, see for example Advanced Organic Chemistry, by Jerry March,4^(th) Edition, Wiley Interscience, pages 131-133, 1992. Examples ofbridged ring systems include bicyclo[2.2.1]heptane,aza-bicyclo[2.2.1]heptane, bicyclo[2.2.2]octane,aza-bicyclo[2.2.2]octane, bicyclo[3.2.1]octane andaza-bicyclo[3.2.1]octane. A particular example of a bridged ring systemis the 1-aza-bicyclo[2.2.2]octan-3-yl group.

Where reference is made herein to carbocyclic and heterocyclic groups,the carbocyclic or heterocyclic ring can, unless the context indicatesotherwise, be unsubstituted or substituted by one or more substituentgroups R¹⁰ selected from halogen, hydroxy, trifluoromethyl, cyano,nitro, carboxy, amino, mono- or di-C₁₋₄ hydrocarbylamino, carbocyclicand heterocyclic groups having from 3 to 12 ring members; a groupR^(a)-R^(b) wherein R^(a) is a bond, O, CO, X¹C(X²), C(X²)X¹, X¹C(X²)X¹,S, SO, SO₂, NR^(c), SO₂NR^(c) or NR^(c)SO₂; and R^(b) is selected fromhydrogen, carbocyclic and heterocyclic groups having from 3 to 12 ringmembers, and a C₁₋₈ hydrocarbyl group optionally substituted by one ormore substituents selected from hydroxy, oxo, halogen, cyano, nitro,carboxy, amino, mono- or di-C₁₋₄ hydrocarbylamino, carbocyclic andheterocyclic groups having from 3 to 12 ring members and wherein one ormore carbon atoms of the C₁₋₈ hydrocarbyl group may optionally bereplaced by O, S, SO, SO₂, NR^(c), X¹C(X²), C(X²)X¹ or X¹C(X²)X¹;

-   -   R^(c) is selected from hydrogen and C₁₋₄ hydrocarbyl; and    -   X¹ is O, S or NR^(c) and X² is ═O, ═S or ═NR^(c).

Where the substituent group R¹⁰ comprises or includes a carbocyclic orheterocyclic group, the said carbocyclic or heterocyclic group may beunsubstituted or may itself be substituted with one or more furthersubstituent groups R¹⁰. In one sub-group of compounds of the formula(I), such further substituent groups R¹⁰ may include carbocyclic orheterocyclic groups, which are typically not themselves furthersubstituted. In another sub-group of compounds of the formula (I), thesaid further substituents do not include carbocyclic or heterocyclicgroups but are otherwise selected from the groups listed above in thedefinition of R¹⁰.

The substituents R¹⁰ may be selected such that they contain no more than20 non-hydrogen atoms, for example, no more than 15 non-hydrogen atoms,e.g. no more than 12, or 11, or 10, or 9, or 8, or 7, or 6, or 5non-hydrogen atoms.

Where the carbocyclic and heterocyclic groups have a pair ofsubstituents on adjacent ring atoms, the two substituents may be linkedso as to form a cyclic group. Thus, two adjacent groups R¹⁰, togetherwith the carbon atoms or heteroatoms to which they are attached may forma 5-membered heteroaryl ring or a 5- or 6-membered non-aromaticcarbocyclic or heterocyclic ring, wherein the said heteroaryl andheterocyclic groups contain up to 3 heteroatom ring members selectedfrom N, O and S. For example, an adjacent pair of substituents onadjacent carbon atoms of a ring may be linked via one or moreheteroatoms and optionally substituted alkylene groups to form a fusedoxa-, dioxa-, aza-, diaza- or oxa-aza-cycloalkyl group.

Examples of such linked substituent groups include:

Examples of halogen substituents include fluorine, chlorine, bromine andiodine. Fluorine and chlorine are particularly preferred.

In the definition of the compounds of the formula (I) above and as usedhereinafter, the term “hydrocarbyl” is a generic term encompassingaliphatic, alicyclic and aromatic groups having an all-carbon backboneand consisting of carbon and hydrogen atoms, except where otherwisestated.

In certain cases, as defined herein, one or more of the carbon atomsmaking up the carbon backbone may be replaced by a specified atom orgroup of atoms.

Examples of hydrocarbyl groups include alkyl, cycloalkyl, cycloalkenyl,carbocyclic aryl, alkenyl, alkynyl, cycloalkylalkyl, cycloalkenylalkyl,and carbocyclic aralkyl, aralkenyl and aralkynyl groups. Such groups canbe unsubstituted or, where stated, substituted by one or moresubstituents as defined herein. The examples and preferences expressedbelow apply to each of the hydrocarbyl substituent groups orhydrocarbyl-containing substituent groups referred to in the variousdefinitions of substituents for compounds of the formula (I) unless thecontext indicates otherwise.

Preferred non-aromatic hydrocarbyl groups are saturated groups such asalkyl and cycloalkyl groups.

Generally by way of example, the hydrocarbyl groups can have up to eightcarbon atoms, unless the context requires otherwise. Within the sub-setof hydrocarbyl groups having 1 to 8 carbon atoms, particular examplesare C₁₋₆ hydrocarbyl groups, such as C₁₋₄ hydrocarbyl groups (e.g. C₁₋₃hydrocarbyl groups or C₁₋₂ hydrocarbyl groups), specific examples beingany individual value or combination of values selected from C₁, C₂, C₃,C₄, C₅, C₆, C₇ and C₈ hydrocarbyl groups.

The term “alkyl” covers both straight chain and branched chain alkylgroups. Examples of alkyl groups include methyl, ethyl, propyl,isopropyl, n-butyl, isobutyl, tert-butyl, n-pentyl, 2-pentyl, 3-pentyl,2-methyl butyl, 3-methyl butyl, and n-hexyl and its isomers. Within thesub-set of alkyl groups having 1 to 8 carbon atoms, particular examplesare C₁₋₆ alkyl groups, such as C₁₋₄ alkyl groups (e.g. C₁₋₃ alkyl groupsor C₁₋₂ alkyl groups).

Examples of cycloalkyl groups are those derived from cyclopropane,cyclobutane, cyclopentane, cyclohexane and cycloheptane. Within thesub-set of cycloalkyl groups the cycloalkyl group will have from 3 to 8carbon atoms, particular examples being C₃₋₆ cycloalkyl groups.

Examples of alkenyl groups include, but are not limited to, ethenyl(vinyl), 1-propenyl, 2-propenyl (allyl), isopropenyl, butenyl,buta-1,4-dienyl, pentenyl, and hexenyl. Within the sub-set of alkenylgroups the alkenyl group will have 2 to 8 carbon atoms, particularexamples being C₂₋₆ alkenyl groups, such as C₂₋₄ alkenyl groups.

Examples of cycloalkenyl groups include, but are not limited to,cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl andcyclohexenyl. Within the sub-set of cycloalkenyl groups the cycloalkenylgroups have from 3 to 8 carbon atoms, and particular examples are C₃₋₆cycloalkenyl groups.

Examples of alkynyl groups include, but are not limited to, ethynyl and2-propynyl (propargyl) groups. Within the sub-set of alkynyl groupshaving 2 to 8 carbon atoms, particular examples are C₂₋₆ alkynyl groups,such as C₂₋₄ alkynyl groups.

Examples of carbocyclic aryl groups include substituted andunsubstituted phenyl groups.

Examples of cycloalkylalkyl, cycloalkenylalkyl, carbocyclic aralkyl,aralkenyl and aralkynyl groups include phenethyl, benzyl, styryl,phenylethynyl, cyclohexylmethyl, cyclopentylmethyl, cyclobutylmethyl,cyclopropylmethyl and cyclopentenylmethyl groups.

When present, and where stated, a hydrocarbyl group can be optionallysubstituted by one or more substituents selected from hydroxy, oxo,alkoxy, carboxy, halogen, cyano, nitro, amino, mono- or di-C₁₋₄hydrocarbylamino, and monocyclic or bicyclic carbocyclic andheterocyclic groups having from 3 to 12 (typically 3 to 10 and moreusually 5 to 10) ring members. Preferred substituents include halogensuch as fluorine. Thus, for example, the substituted hydrocarbyl groupcan be a partially fluorinated or perfluorinated group such asdifluoromethyl or trifluoromethyl. In one embodiment preferredsubstituents include monocyclic carbocyclic and heterocyclic groupshaving 3-7 ring members, more usually 3, 4, 5 or 6 ring members.

Where stated, one or more carbon atoms of a hydrocarbyl group mayoptionally be replaced by O, S, SO, SO₂, NR^(c), X¹C(X²), C(X²)X¹ orX¹C(X²)X¹ (or a sub-group thereof) wherein X¹ and X² are as hereinbeforedefined, provided that at least one carbon atom of the hydrocarbyl groupremains. For example, 1, 2, 3 or 4 carbon atoms of the hydrocarbyl groupmay be replaced by one of the atoms or groups listed, and the replacingatoms or groups may be the same or different. In general, the number oflinear or backbone carbon atoms replaced will correspond to the numberof linear or backbone atoms in the group replacing them. Examples ofgroups in which one or more carbon atom of the hydrocarbyl group havebeen replaced by a replacement atom or group as defined above includeethers and thioethers (C replaced by O or S), amides, esters, thioamidesand thioesters (C—C replaced by X¹C(X²) or C(X²)X¹), sulphones andsulphoxides (C replaced by SO or SO₂), amines (C replaced by NR^(c)).Further examples include ureas, carbonates and carbamates (C—C—Creplaced by X¹C(X²)X¹).

Where an amino group has two hydrocarbyl substituents, they may,together with the nitrogen atom to which they are attached, andoptionally with another heteroatom such as nitrogen, sulphur, or oxygen,link to form a ring structure of 4 to 7 ring members, more usually 5 to6 ring members.

The term “aza-cycloalkyl” as used herein refers to a cycloalkyl group inwhich one of the carbon ring members has been replaced by a nitrogenatom. Thus examples of aza-cycloalkyl groups include piperidine andpyrrolidine. The term “oxa-cycloalkyl” as used herein refers to acycloalkyl group in which one of the carbon ring members has beenreplaced by an oxygen atom. Thus examples of oxa-cycloalkyl groupsinclude tetrahydrofuran and tetrahydropyran. In an analogous manner, theterms “diaza-cycloalkyl”, “dioxa-cycloalkyl” and “aza-oxa-cycloalkyl”refer respectively to cycloalkyl groups in which two carbon ring membershave been replaced by two nitrogen atoms, or by two oxygen atoms, or byone nitrogen atom and one oxygen atom.

The definition “R^(a)-R^(b)” as used herein, either with regard tosubstituents present on a carbocyclic or heterocyclic moiety, or withregard to other substituents present at other locations on the compoundsof the formula (I), includes inter alia compounds wherein R^(a) isselected from a bond, O, CO, OC(O), SC(O), NR^(c)C(O), OC(S), SC(S),NR^(c)C(S), OC(NR^(c)), SC(NR^(c)), NR^(c)C(NR^(c)), C(O)O, C(O)S,C(O)NR^(c), C(S)O, C(S)S, C(S)NR^(c), C(NR^(c))O, C(NR^(c))S,C(NR^(c))NR^(c), OC(O)O, SC(O)O, NR^(c)C(O)O, OC(S)O, SC(S)O,NR^(c)C(S)O, OC(NR^(c))O, SC(NR^(c))O, NR^(c)C(NR^(c))O, OC(O)S, SC(O)S,NR^(c)C(O)S, OC(S)S, SC(S)S, NR^(c)C(S)S, OC(NR^(c))S, SC(NR^(c))S,NR^(c)C(NR^(c))S, OC(O)NR^(c), SC(O)NR^(c), NR^(c)C(O)NR^(c),OC(S)NR^(c), SC(S)NR^(c), NR^(c)C(S)NR^(c), OC(NR^(c))NR^(c),SC(NR^(c))NR^(c), NR^(c)C(NR^(c)NR^(c), S, SO, SO₂, NR^(c), SO₂NR^(c)and NR^(c)SO₂ wherein R^(c) is as hereinbefore defined.

The moiety R^(b) can be hydrogen or it can be a group selected fromcarbocyclic and heterocyclic groups having from 3 to 12 ring members(typically 3 to 10 and more usually from 5 to 10), and a C₁₋₈hydrocarbyl group optionally substituted as hereinbefore defined.Examples of hydrocarbyl, carbocyclic and heterocyclic groups are as setout above.

When R^(a) is O and R^(b) is a C₁₋₈ hydrocarbyl group, R^(a) and R^(b)together form a hydrocarbyloxy group. Preferred hydrocarbyloxy groupsinclude saturated hydrocarbyloxy such as alkoxy (e.g. C₁₋₆ alkoxy, moreusually C₁₋₄ alkoxy such as ethoxy and methoxy, particularly methoxy),cycloalkoxy (e.g. C₃₋₆ cycloalkoxy such as cyclopropyloxy,cyclobutyloxy, cyclopentyloxy and cyclohexyloxy) and cycloalkyalkoxy(e.g. C₃₋₆ cycloalkyl-C₁₋₂ alkoxy such as cyclopropylmethoxy).

The hydrocarbyloxy groups can be substituted by various substituents asdefined herein. For example, the alkoxy groups can be substituted byhalogen (e.g. as in difluoromethoxy and trifluoromethoxy), hydroxy (e.g.as in hydroxyethoxy), C₁₋₂ alkoxy (e.g. as in methoxyethoxy),hydroxy-C₁₋₂ alkyl (as in hydroxyethoxyethoxy) or a cyclic group (e.g. acycloalkyl group or non-aromatic heterocyclic group as hereinbeforedefined). Examples of alkoxy groups bearing a non-aromatic heterocyclicgroup as a substituent are those in which the heterocyclic group is asaturated cyclic amine such as morpholine, piperidine, pyrrolidine,piperazine, C₁₋₄-alkyl-piperazines, C₃₋₇-cycloalkyl-piperazines,tetrahydropyran or tetrahydrofuran and the alkoxy group is a C₁₋₄ alkoxygroup, more typically a C₁₋₃ alkoxy group such as methoxy, ethoxy orn-propoxy.

Alkoxy groups substituted by a monocyclic group such as pyrrolidine,piperidine, morpholine and piperazine and N-substituted derivativesthereof such as N-benzyl, N—C₁₋₄ acyl and N—C₁₋₄ alkoxycarbonyl.Particular examples include pyrrolidinoethoxy, piperidinoethoxy andpiperazinoethoxy.

When R^(a) is a bond and R^(b) is a C₁₋₈ hydrocarbyl group, examples ofhydrocarbyl groups R^(a)-R^(b) are as hereinbefore defined. Thehydrocarbyl groups may be saturated groups such as cycloalkyl and alkyland particular examples of such groups include methyl, ethyl andcyclopropyl. The hydrocarbyl (e.g. alkyl) groups can be substituted byvarious groups and atoms as defined herein. Examples of substitutedalkyl groups include alkyl groups substituted by one or more halogenatoms such as fluorine and chlorine (particular examples includingbromoethyl, chloroethyl and trifluoromethyl), or hydroxy (e.g.hydroxymethyl and hydroxyethyl), C₁₋₈ acyloxy (e.g. acetoxymethyl andbenzyloxymethyl), amino and mono- and dialkylamino (e.g. aminoethyl,methylaminoethyl, dimethylaminomethyl, dimethylaminoethyl andtert-butylaminomethyl), alkoxy (e.g. C₁₋₂ alkoxy such as methoxy—as inmethoxyethyl), and cyclic groups such as cycloalkyl groups, aryl groups,heteroaryl groups and non-aromatic heterocyclic groups as hereinbeforedefined).

Particular examples of alkyl groups substituted by a cyclic group arethose wherein the cyclic group is a saturated cyclic amine such asmorpholine, piperidine, pyrrolidine, piperazine, C₁₋₄-alkyl-piperazines,C₃₋₇-cycloalkyl-piperazines, tetrahydropyran or tetrahydrofuran and thealkyl group is a C₁₋₄ alkyl group, more typically a C₁₋₃ alkyl groupsuch as methyl, ethyl or n-propyl. Specific examples of alkyl groupssubstituted by a cyclic group include pyrrolidinomethyl,pyrrolidinopropyl, morpholinomethyl, morpholinoethyl, morpholinopropyl,piperidinylmethyl, piperazinomethyl and N-substituted forms thereof asdefined herein.

Particular examples of alkyl groups substituted by aryl groups andheteroaryl groups include benzyl and pyridylmethyl groups.

When R^(a) is SO₂NR^(c), R^(b) can be, for example, hydrogen or anoptionally substituted C₁₋₈ hydrocarbyl group, or a carbocyclic orheterocyclic group. Examples of R^(a)-R^(b) where R^(a) is SO₂NR^(c)include aminosulphonyl, C₁₋₄alkylaminosulphonyl and di-C₁₋₄alkylaminosulphonyl groups, and sulphonamides formed from a cyclic aminogroup such as piperidine, morpholine, pyrrolidine, or an optionallyN-substituted piperazine such as N-methyl piperazine.

Examples of groups R^(a)-R^(b) where R^(a) is SO₂ includealkylsulphonyl, heteroarylsulphonyl and arylsulphonyl groups,particularly monocyclic aryl and heteroaryl sulphonyl groups. Particularexamples include methylsulphonyl, phenylsulphonyl and toluenesulphonyl.

When R^(a) is NR^(c), R^(b) can be, for example, hydrogen or anoptionally substituted C₁₋₈ hydrocarbyl group, or a carbocyclic orheterocyclic group. Examples of R^(a)-R^(b) where R^(a) is NR^(c)include amino, C₁₋₄ alkylamino (e.g. methylamino, ethylamino,propylamino, isopropylamino, tert-butylamino), di-C₁₋₄ alkylamino (e.g.dimethylamino and diethylamino) and cycloalkylamino (e.g.cyclopropylamino, cyclopentylamino and cyclohexylamino).

Specific Embodiments of and Preferences for X, Y, A, R^(g), R¹ to R⁴ andR¹⁰ X

In formula (I), X is a group R¹-A-NR⁴— or a 5- or 6-membered carbocyclicor heterocyclic ring.

In one embodiment, X is a group R¹-A-NR⁴—.

In another embodiment, X is a 5- or 6-membered carbocyclic orheterocyclic ring.

A

In formula (I), A is a bond, C═O, NR^(g)(C═O) or O(C═O). It will beappreciated that the moiety R¹-A-NR⁴ linked to the 4-position of thepyrazole ring can therefore take the form of an amine R¹—NR⁴, an amideR¹—C(═O)NR⁴, a urea R¹—NR^(g)C(═O)NR⁴ or a carbamate R¹—OC(═O)NR⁴.

In one preferred group of compounds of the invention, A is C═O and hencethe group R¹-A-NR⁴ takes the form of an amide R¹—C(═O)NR⁴. In anothergroup of compounds of the invention, A is a bond and hence the groupR¹-A-NR⁴ takes the form of an amine R¹—NR⁴.

R⁴

R⁴ is hydrogen or a C₁₋₄ hydrocarbyl group optionally substituted byhalogen (e.g. fluorine), hydroxyl or C₁₋₄ alkoxy (e.g. methoxy).

The number of optional substitutents on the hydrocarbyl group typicallywill vary according to the nature of the substituent. For example, wherethe substituent is halogen, there may be from one to three halogen atomspresent, preferably two or three. Where the substituent is hydroxyl oran alkoxy group, typically there will be only a single such substituentpresent

R⁴ is preferably hydrogen or C₁₋₃ alkyl, more preferably hydrogen ormethyl and most preferably is hydrogen.

R^(g)

R^(g) is hydrogen or a C₁₋₄ hydrocarbyl group optionally substituted byhydroxyl or C₁₋₄ alkoxy (e.g. methoxy).

When R^(g) is C₁₋₄ hydrocarbyl substituted by hydroxyl or C₁₋₄ alkoxy,typically there is only one such substituent present.

Preferably R^(g) is hydrogen or C₁₋₃ alkyl, more preferably hydrogen ormethyl and most preferably R^(g) is hydrogen.

R²

R² is hydrogen, halogen, C₁₋₄ alkoxy, or a C₁₋₄ hydrocarbyl groupoptionally substituted by halogen, hydroxyl or C₁₋₄ alkoxy.

When R² is halogen, preferably it is selected from chlorine and fluorineand more preferably it is fluorine.

When R² is C₁₋₄ alkoxy, it can be, for example, C₁₋₃ alkoxy, morepreferably C₁₋₂ alkoxy and most preferably methoxy.

When R² is an optionally substituted C₁₋₄ hydrocarbyl group, thehydrocarbyl group is preferably a C₁₋₃ hydrocarbyl group, morepreferably a C₁₋₂ hydrocarbyl group, for example an optionallysubstituted methyl group. The optional substituents for the optionallysubstituted hydrocarbyl group are preferably selected from fluorine,hydroxyl and methoxy.

The number of optional substituents on the hydrocarbyl group typicallywill vary according to the nature of the substituent. For example, wherethe substituent is halogen, there may be from one to three halogen atomspresent, preferably two or three. Where the substituent is hydroxyl ormethoxy, typically there will be only a single such substituent present.

The hydrocarbyl groups constituting R² are preferably saturatedhydrocarbyl groups. Examples of saturated hydrocarbyl groups includemethyl, ethyl, n-propyl, i-propyl and cyclopropyl.

In one embodiment, R² is hydrogen, halogen, C₁₋₄ alkoxy, or a C₁₋₄hydrocarbyl group optionally substituted by halogen, hydroxyl or C₁₋₄alkoxy.

In another embodiment, R² is hydrogen, fluorine, chlorine, methoxy, or aC₁₋₃ hydrocarbyl group optionally substituted by fluorine, hydroxyl ormethoxy.

In a preferred embodiment, R² is hydrogen or methyl, most preferablyhydrogen.

R¹

R¹ is hydrogen, a carbocyclic or heterocyclic group having from 3 to 12ring members, or a C₁₋₈ hydrocarbyl group optionally substituted by oneor more substituents selected from halogen (e.g. fluorine), hydroxy,C₁₋₄ hydrocarbyloxy, amino, mono- or di-C₁₋₄ hydrocarbylamino, andcarbocyclic or heterocyclic groups having from 3 to 12 ring members, andwherein 1 or 2 of the carbon atoms of the hydrocarbyl group mayoptionally be replaced by an atom or group selected from O, S, NH, SO,SO₂. Examples of carbocyclic or heterocyclic groups and hydrocarbylgroups and general preferences for such groups are as set out above inthe General Preferences and Definitions section, and as set out below.

In one embodiment, R¹ is an aryl or heteroaryl group.

When R¹ is a heteroaryl group, particular heteroaryl groups includemonocyclic heteroaryl groups containing up to three heteroatom ringmembers selected from O, S and N, and bicyclic heteroaryl groupscontaining up to 2 heteroatom ring members selected from O, S and N andwherein both rings are aromatic.

Examples of such groups include furanyl (e.g. 2-furanyl or 3-furanyl),indolyl (e.g. 3-indolyl, 6-indolyl), 2,3-dihydro-benzo[1,4]dioxinyl(e.g. 2,3-dihydro-benzo[1,4]dioxin-5-yl), pyrazolyl (e.g.pyrazole-5-yl), pyrazolo[1,5-a]pyridinyl (e.g.pyrazolo[1,5-a]pyridine-3-yl), oxazolyl (e.g.), isoxazolyl (e.g.isoxazol-4-yl), pyridyl (e.g. 2-pyridyl, 3-pyridyl, 4-pyridyl),quinolinyl (e.g. 2-quinolinyl), pyrrolyl (e.g. 3-pyrrolyl), imidazolyland thienyl (e.g. 2-thienyl, 3-thienyl).

One sub-group of heteroaryl groups R¹ consists of furanyl (e.g.2-furanyl or 3-furanyl), indolyl, oxazolyl, isoxazolyl, pyridyl,quinolinyl, pyrrolyl, imidazolyl and thienyl.

A preferred sub-set of R¹ heteroaryl groups includes 2-furanyl,3-furanyl, pyrrolyl, imidazolyl and thienyl.

Preferred aryl groups R¹ are phenyl groups.

The group R¹ can be an unsubstituted or substituted carbocylic orheterocyclic group in which one or more substituents can be selectedfrom the group R¹⁰ as hereinbefore defined. In one embodiment, thesubstituents on R¹ may be selected from the group R^(10a) consisting ofhalogen, hydroxy, trifluoromethyl, cyano, nitro, carboxy, a groupR^(a)-R^(b) wherein R^(a) is a bond, O, CO, X³C(X⁴), C(X⁴)X³, X³C(X⁴)X³,S, SO, or SO₂, and R^(b) is selected from hydrogen and a C₁₋₈hydrocarbyl group optionally substituted by one or more substituentsselected from hydroxy, oxo, halogen, cyano, nitro, carboxy andmonocyclic non-aromatic carbocyclic or heterocyclic groups having from 3to 6 ring members; wherein one or more carbon atoms of the C₁₋₈hydrocarbyl group may optionally be replaced by O, S, SO, SO₂, X³C(X⁴),C(X⁴)X³ or X³C(X⁴)X³; X³ is O or S; and X⁴ is ═O or ═S.

Where the carbocyclic and heterocyclic groups have a pair ofsubstituents on adjacent ring atoms, the two substituents may be linkedso as to form a cyclic group. Thus, two adjacent groups R¹⁰, togetherwith the carbon atoms or heteroatoms to which they are attached may forma 5-membered heteroaryl ring or a 5- or 6-membered non-aromaticcarbocyclic or heterocyclic ring, wherein the said heteroaryl andheterocyclic groups contain up to 3 heteroatom ring members selectedfrom N, O and S. In particular the two adjacent groups R¹⁰, togetherwith the carbon atoms or heteroatoms to which they are attached, mayform a 6-membered non-aromatic heterocyclic ring, containing up to 3, inparticular 2, heteroatom ring members selected from N, O and S. Moreparticularly the two adjacent groups R¹⁰ may form a 6-memberednon-aromatic heterocyclic ring, containing 2 heteroatom ring membersselected from N, or O, such as dioxan e.g. [1,4 dioxan]. In oneembodiment R¹ is a carbocyclic group e.g. phenyl having a pair ofsubstituents on adjacent ring atoms linked so as to form a cyclic groupe.g. to form 2,3-dihydro-benzo[1,4]dioxine.

More particularly, the substituents on R¹ may be selected from halogen,hydroxy, trifluoromethyl, a group R^(a)-R^(b) wherein R^(a) is a bond orO, and R^(b) is selected from hydrogen and a C₁₋₄ hydrocarbyl groupoptionally substituted by one or more substituents selected fromhydroxyl, halogen (preferably fluorine) and 5 and 6 membered saturatedcarbocyclic and heterocyclic groups (for example groups containing up totwo heteroatoms selected from O, S and N, such as unsubstitutedpiperidine, pyrrolidino, morpholino, piperazino and N-methylpiperazino).

The group R¹ may be substituted by more than one substituent. Thus, forexample, there may be 1 or 2 or 3 or 4 substituents. In one embodiment,where R¹ is a six membered ring (e.g. a carbocyclic ring such as aphenyl ring), there may be one, two or three substituents and these maybe located at the 2-, 3-, 4- or 6-positions around the ring. By way ofexample, a phenyl group R¹ may be 2-monosubstituted, 3-monosubstituted,2,6-disubstituted, 2,3-disubstituted, 2,4-disubstituted2,5-disubstituted, 2,3,6-trisubstituted or 2,4,6-trisubstituted. Moreparticularly, a phenyl group R¹ may be monosubstituted at the 2-positionor disubstituted at positions 2- and 6- with substituents selected fromfluorine, chlorine and R^(a)-R^(b), where R^(a) is O and R^(b) is C₁₋₄alkyl (e.g. methyl or ethyl). In one embodiment, fluorine is a preferredsubstituent. In another embodiment, preferred substituents are selectedfrom fluorine, chlorine and methoxy.

Particular examples of non-aromatic groups R¹ include unsubstituted orsubstituted (by one or more groups R¹⁰) monocyclic cycloalkyl groups.Examples of such cycloalkyl groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl and cycloheptyl; more typically cyclopropyl,cyclobutyl, cyclopentyl and cyclohexyl, particularly cyclohexyl.

Further examples of non-aromatic groups R¹ include unsubstituted orsubstituted (by one or more groups R¹⁰) heterocyclic groups having from3 to 12 ring members, typically 4 to 12 ring members, and more usuallyfrom 5 to 10 ring members. Such groups can be monocyclic or bicyclic,for example, and typically have from 1 to 5 heteroatom ring members(more usually 1,2, 3 or 4 heteroatom ring members) typically selectedfrom nitrogen, oxygen and sulphur.

When sulphur is present, it may, where the nature of the adjacent atomsand groups permits, exist as —S—, —S(O)— or —S(O)₂—.

The heterocylic groups can contain, for example, cyclic ether moieties(e.g as in tetrahydrofuran and dioxane), cyclic thioether moieties (e.g.as in tetrahydrothiophene and dithiane), cyclic amine moieties (e.g. asin pyrrolidine), cyclic amides (e.g. as in pyrrolidone), cyclic esters(e.g. as in butyrolactone), cyclic thioamides and thioesters, cyclicsulphones (e.g. as in sulpholane and sulpholene), cyclic sulphoxides,cyclic sulphonamides and combinations thereof (e.g. morpholine andthiomorpholine and its S-oxide and S,S-dioxide).

In one sub-set of heterocyclic groups R¹, the heterocyclic groupscontain cyclic ether moieties (e.g as in tetrahydrofuran and dioxane),cyclic thioether moieties (e.g. as in tetrahydrothiophene and dithiane),cyclic amine moieties (e.g. as in pyrrolidine), cyclic sulphones (e.g.as in sulpholane and sulpholene), cyclic sulphoxides, cyclicsulphonamides and combinations thereof (e.g. thiomorpholine).

Examples of monocyclic non-aromatic heterocyclic groups R¹ include 5-,6- and 7-membered monocyclic heterocyclic groups such as morpholine,piperidine (e.g. 1-piperidinyl, 2-piperidinyl 3-piperidinyl and4-piperidinyl), pyrrolidine (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl and3-pyrrolidinyl), pyrrolidone, pyran (2H-pyran or 4H-pyran),dihydrothiophene, dihydropyran, dihydrofuran, dihydrothiazole,tetrahydrofuran, tetrahydrothiophene, dioxane, tetrahydropyran (e.g.4-tetrahydro pyranyl), imidazoline, imidazolidinone, oxazoline,thiazoline, 2-pyrazoline, pyrazolidine, piperazine, and N-alkylpiperazines such as N-methyl piperazine. Further examples includethiomorpholine and its S-oxide and S,S-dioxide (particularlythiomorpholine). Still further examples include N-alkyl piperidines suchas N-methyl piperidine.

One sub-group of non-aromatic heterocyclic groups R¹ includesunsubstituted or substituted (by one or more groups R¹⁰) 5-, 6- and7-membered monocyclic heterocyclic groups such as morpholine, piperidine(e.g. 1-piperidinyl, 2-piperidinyl 3-piperidinyl and 4-piperidinyl),pyrrolidine (e.g. 1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl),pyrrolidone, piperazine, and N-alkyl piperazines such as N-methylpiperazine, wherein a particular sub-set consists of pyrrolidine,piperidine, morpholine, thiomorpholine and N-methyl piperazine.

In general, preferred non-aromatic heterocyclic groups includepyrrolidine, piperidine, morpholine, thiomorpholine, thiomorpholineS,S-dioxide, piperazine, N-alkyl piperazines, and N-alkyl piperidines.

Another particular sub-set of heterocyclic groups consists ofpyrrolidine, piperidine, morpholine and N-alkyl piperazines, andoptionally, N-methyl piperazine and thiomorpholine.

When R¹ is a C₁₋₈ hydrocarbyl group substituted by a carbocyclic orheterocyclic group, the carbocyclic and heterocyclic groups can bearomatic or non-aromatic and can be selected from the examples of suchgroups set out hereinabove. The substituted hydrocarbyl group istypically a saturated C₁ hydrocarbyl group such as an alkyl group,preferably a CH₂ or CH₂CH₂ group. Where the substituted hydrocarbylgroup is a C₂₋₄ hydrocarbyl group, one of the carbon atoms and itsassociated hydrogen atoms may be replaced by a sulphonyl group; forexample as in the moiety SO₂CH₂.

When the carbocyclic or heterocylic group attached to the a C₁₋₈hydrocarbyl group is aromatic, examples of such groups includemonocyclic aryl groups and monocyclic heteroaryl groups containing up tofour heteroatom ring members selected from O, S and N, and bicyclicheteroaryl groups containing up to 2 heteroatom ring members selectedfrom O, S and N and wherein both rings are aromatic.

Examples of such groups are set out in the “General Preferences andDefinitions” section above.

Particular examples of such groups include furanyl (e.g. 2-furanyl or3-furanyl), indolyl, oxazolyl, isoxazolyl, pyridyl, quinolinyl,pyrrolyl, imidazolyl and thienyl. Particular examples of aryl andheteroaryl groups as substituents for a C₁₋₈ hydrocarbyl group includephenyl, imidazolyl, tetrazolyl, triazolyl, indolyl, 2-furanyl,3-furanyl, pyrrolyl and thienyl. Such groups may be substituted by oneor more substituents R¹⁰ or R^(10a) as defined herein.

When R¹ is a C₁₋₈ hydrocarbyl group substituted by a non-aromaticcarbocyclic or heterocyclic group, the non-aromatic or heterocyclicgroup may be a group selected from the lists of such groups set outhereinabove. For example, the non-aromatic group can be a monocyclicgroup having from 4 to 7 ring members, e.g. 5 to 7 ring members, andtypically containing from 0 to 3, more typically 0, 1 or 2, heteroatomring members selected from O, S and N. When the cyclic group is acarbocyclic group, it may additionally be selected from monocyclicgroups having 3 ring members. Particular examples include monocycliccycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl and cycloheptyl, and 5-, 6- and 7-membered monocyclicheterocyclic groups such as morpholine, piperidine (e.g. 1-piperidinyl,2-piperidinyl, 3-piperidinyl and 4-piperidinyl), pyrrolidine (e.g.1-pyrrolidinyl, 2-pyrrolidinyl and 3-pyrrolidinyl), pyrrolidone,piperazine, and N-alkyl piperazines such as N-methyl piperazine. Ingeneral, preferred non-aromatic heterocyclic groups include pyrrolidine,piperidine, morpholine, thiomorpholine and N-methyl piperazine.

When R¹ is an optionally substituted C₁₋₈ hydrocarbyl group, thehydrocarbyl group may be as hereinbefore defined, and is preferably upto four carbon atoms in length, more usually up to three carbon atoms inlength for example one or two carbon atoms in length.

In one embodiment, the hydrocarbyl group is saturated and may be acyclicor cyclic, for example acyclic. An acyclic saturated hydrocarbyl group(i.e. an alkyl group) may be a straight chain or branched alkyl group.

Examples of straight chain alkyl groups R¹ include methyl, ethyl, propyland butyl.

Examples of branched chain alkyl groups R¹ include isopropyl, isobutyl,tert-butyl and 2,2-dimethylpropyl.

In one embodiment, the hydrocarbyl group is a linear saturated grouphaving from 1-6 carbon atoms, more usually 1-4 carbon atoms, for example1-3 carbon atoms, e.g. 1, 2 or 3 carbon atoms. When the hydrocarbylgroup is substituted, particular examples of such groups are substituted(e.g. by a carbocyclic or heterocyclic group) methyl and ethyl groups.

A C₁₋₈ hydrocarbyl group R¹ can be optionally substituted by one or moresubstituents selected from halogen (e.g. fluorine), hydroxy, C₁₋₄hydrocarbyloxy, amino, mono- or di-C₁₋₄ hydrocarbylamino, andcarbocyclic or heterocyclic groups having from 3 to 12 ring members, andwherein 1 or 2 of the carbon atoms of the hydrocarbyl group mayoptionally be replaced by an atom or group selected from O, S, NH, SO,SO₂. Particular substituents for the hydrocarbyl group include hydroxy,chlorine, fluorine (e.g. as in trifluoromethyl), methoxy, ethoxy, amino,methylamino and dimethylamino, preferred substituents being hydroxy andfluorine.

When A is C═O, particular groups R¹—CO are the groups set out in Table 1below.

In Table 1, the point of attachment of the group to the nitrogen atom ofthe pyrazole-4-amino group is represented by the terminal single bondextending from the carbonyl group. Thus, by way of illustration, group Bin the table is the trifluoroacetyl group, group D in the table is thephenylacetyl group and group I in the table is the3-(4-chlorophenyl)propionyl group.

TABLE 1 Examples of the group R¹—CO CH₃—C(═O)— A CF₃—C(═O)— B

  C

  D

  E

  F

  G

  H

  I

  J

  K

  L

  M

  N

  O

  P

  Q

  R

  S

  T

  U

  V

  W

  X

  Y

  Z

  AA

  AB

  AC

  AD

  AE

  AF

  AG

  AH

  AI

  AJ

  AK

  AL

  AM

  AN

  AO

  AP

  AQ

  AR

  AS

  AT

  AU

  AV

  AW

  AX

  AY

  AZ

  BA

  BB

  BC

  BD

  BE

  BF

  BG

  BH

  BI

  BJ

  BK

  BL

  BM

  BN

  BO

  BP

  BQ

  BR

  BS

  BT

  BU

  BV

  BW

  BX

  BY

  BZ

  BAA

  BAB

  BAC

  BAD

  BAE

  BAF

  BAG

  BAH

  BAI

  BAJ

  BAK

  BAL

  BAM

  BAN

  BAO

One sub-group of groups R¹—CO consists of groups A to BF in Table 1above.

Another sub-group of groups R¹—CO consists of groups A to BS in Table 1above.

One set of preferred groups R¹—CO consists of the groups J, AB, AH, AJ,AL, AS, AX, AY, AZ, BA, BB, BD, BH, BL, BQ, BS and BAI

Another set of preferred groups R¹—CO consists of the groups J, AB, AH,AJ, AL, AS, AX, AY, AZ, BA, BB, BD, BH, BL, BQ and BS.

More preferred groups R¹—CO— are AJ, AX, BQ, BS and BAI.

One particularly preferred sub-set of groups R¹—CO— consists of AJ, BQand BS.

Another particularly preferred sub-set of groups R¹—CO— consists of AJand BQ.

When X is R¹-A-NR⁴ and A is C═O, and R¹ is a phenyl ring bearing asubstituent at the 4-position, the substituent at the 4-position ispreferably other than a phenyl group having a group SO₂NH₂ or SO₂Me atthe ortho-position.

In one general embodiment, R¹ may be other than a substituted orunsubstituted tetrahydroquinoline, chroman, chromene, thiochroman,thiochromene, dihydro-naphthalene or tetrahydronaphthalene group. Moreparticularly, R¹ may be other than a substituted or unsubstitutedtetrahydroquinoline, chroman, chromene, thiochroman, thiochromene,dihydro-naphthalene or tetrahydronaphthalene group linked by itsaromatic ring to the moiety A-NR⁴—.

In another general embodiment, when R¹ is a substituted or unsubstitutedphenyl group, the moiety Y—R³ may be other than hydrogen, unsubstitutedC₁₋₁₀ alkyl, unsubstituted C₅₋₁₀ cycloalkyl, unsubstituted phenyl,unsubstituted alkylphenyl or unsubstituted phenyl-C₁₋₁₀ alkyl.

In the context of the group R¹-A-NR⁴—, when R¹ is an optionallysubstituted hydrocarbyl group and the hydrocarbyl group comprises orcontains a substituted or unsubstituted alkene group, it is preferredthat the carbon-carbon double bond of the alkene group is not directlybonded to the group A.

Also in the context of the group R¹-A-NR⁴—, when R¹ is an optionallysubstituted hydrocarbyl group, the hydrocarbyl group may be other thanan alkene group.

In another general embodiment, when Y is a bond, R³ is hydrogen, A is COand R¹ is a substituted phenyl group, each substituent on the phenylgroup may be other than a group CH₂—P(O)R^(x)R^(y) where R^(x) and R^(y)are each selected from alkoxy and phenyl groups.

Y

In the compounds of the formula (I), Y is a bond or an alkylene chain of1, 2 or 3 carbon atoms in length.

The term “alkylene” has its usual meaning and refers to a divalentsaturated acyclic hydrocarbon chain. The hydrocarbon chain may bebranched or unbranched. Where an alkylene chain is branched, it may haveone or more methyl group side chains. Examples of alkylene groupsinclude —CH₂—, —CH₂—CH₂—, —CH₂—CH₂-CH₂—, CH(CH₃)—, —C(CH₃)₂—,—CH₂—CH(CH₃)—, —CH₂—C(CH₃)₂— and —CH(CH₃)—CH(CH₃)—.

In one embodiment, Y is a bond.

In another embodiment, Y is an alkylene chain.

When Y is an alkylene chain, preferably it is unbranched and moreparticularly contains 1 or 2 carbon atoms, preferably 1 carbon atom.Thus preferred groups Y are —CH₂— and —CH₂—CH₂—, a most preferred groupbeing (CH₂)—.

Where Y is a branched chain, preferably it has no more than two methylside chains. For example, it may have a single methyl side chain. In oneembodiment, Y is a group —CH(Me)—.

In one sub-group of compounds, Y is a bond, CH₂, CH₂CH₂ or CH₂CH(CH₃).

R³

The group R³ is selected from hydrogen and carbocyclic and heterocyclicgroups having from 3 to 12 ring members.

In one sub-group of compounds, Y is a bond and R³ is hydrogen.

In another sub-group of compounds Y is an alkylene chain as hereinbeforedefined and R³ is hydrogen.

In a another sub-group of compounds, Y is a bond or an alkylene chain(e.g. a group —(CH₂)—) and R³ is a carbocyclic or heterocyclic group.

In a further sub-group of compounds, Y is a bond and R³ is a carbocyclicor heterocyclic group.

In a still further sub-group of compounds, Y is an alkylene chain (e.g.a group —(CH₂)—) and R³ is a carbocyclic or heterocyclic group.

The carbocyclic and heterocyclic groups R³ can be aryl, heteroaryl,non-aromatic carbocyclic or non-aromatic heterocyclic and examples ofsuch groups are as set out in detail above in the General Preferencesand Definitions section, and as set out below.

Preferred aryl groups R³ are unsubstituted and substituted phenylgroups.

Examples of heteroaryl groups R³ include monocyclic heteroaryl groupscontaining up to three (and more preferably up to two) heteroatom ringmembers selected from O, S and N. Preferred heteroaryl groups includefive membered rings containing one or two heteroatom ring members andsix membered rings containing a single heteroatom ring member, mostpreferably nitrogen. Particular examples of heteroaryl groups includeunsubstituted or substituted pyridyl, imidazole, pyrazole, thiazole,isothiazole, isoxazole, oxazole, furyl and thiophene groups.

Particular heteroaryl groups are unsubstituted and substituted pyridylgroups, e.g. 2-pyridyl, 3-pyridyl and 4-pyridyl groups, especially 3-and 4-pyridyl groups. When the pyridyl groups are substituted, they canbear one or more substituents, typically no more than two, and moreusually one substituent selected, for example, from C₁₋₄ alkyl (e.g.methyl), halogen (e.g. fluorine or chlorine, preferably chlorine), andC₁₋₄ alkoxy (e.g. methoxy). Substituents on the pyridyl group mayfurther be selected from amino, mono-C₁₋₄ alkylamino and di-C₁₋₄alkylamino, particularly amino.

In one embodiment, when R³ is an aryl (e.g. phenyl) or heteroaryl group,the substituents on the carbocyclic or heterocyclic group may beselected from the group R^(10a) consisting of halogen, hydroxy,trifluoromethyl, cyano, monocyclic carbocyclic and heterocyclic groupshaving from 3 to 7 (typically 5 or 6) ring members, and a groupR^(a)-R^(b) wherein R^(a) is a bond, O, CO, X¹C(X²), C(X²)X¹, X¹C(X²)X¹,S, SO, SO₂, NR^(c), SO₂NR^(c) or NR^(c)SO₂; and R^(b) is selected fromhydrogen, a carbocyclic or heterocyclic group with 3-7 ring members anda C₁₋₈ hydrocarbyl group optionally substituted by one or moresubstituents selected from hydroxy, oxo, halogen, cyano, nitro, carboxy,amino, mono- or di-C₁₋₄ hydrocarbylamino, a carbocyclic or heterocyclicgroup with 3-7 ring members and wherein one or more carbon atoms of theC₁₋₈ hydrocarbyl group may optionally be replaced by O, S, SO, SO₂,NR^(c), X¹C(X²), C(X²)X¹ or X¹C(X²)X¹; and R^(c), X¹ and X² are ashereinbefore defined.

Examples of non-aromatic groups R³ include optionally substituted (byR¹⁰ or R^(10a)) cycloalkyl, oxa-cycloalkyl, aza-cycloalkyl,diaza-cycloalkyl, dioxa-cycloalkyl and aza-oxa-cycloalkyl groups.Further examples include C₇₋₁₀ aza-bicycloalkyl groups such as1-aza-bicyclo[2.2.2]octan-3-yl.

Particular examples of such groups include unsubstituted or substitutedcyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran,morpholine, tetrahydrofuran, piperidine and pyrrolidine groups.

One sub-set of non-aromatic groups R³ consists of cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, tetrahydropyran, tetrahydrofuran,piperidine and pyrrolidine groups.

Preferred non-aromatic groups R³ include unsubstituted or substitutedcyclopentyl, cyclohexyl, tetrahydropyran, tetrahydrofuran, piperidineand pyrrolidine groups,

The non-aromatic groups may be unsubstituted or substituted with one ormore groups R¹⁰ or R^(10a) as hereinbefore defined.

Particular substituents for R³ (e.g. (i) when R³ is an aryl orheteroaryl group or (ii) when R³ is a non-aromatic group) are selectedfrom the group R^(10a) consisting of halogen; hydroxy; monocycliccarbocyclic and heterocyclic groups having from 3 to 6 ring members andcontaining up to 2 heteroataom ring members selected from O, N and S;and a group R^(a)-R^(b) wherein R^(a) is a bond, O, CO, CO₂, SO₂, NH,SO₂NH or NHSO₂; and R^(b) is selected from hydrogen, a carbocyclic orheterocyclic group with 3-6 ring members and containing up to 2heteroatom ring members selected from O, N and S; and a C₁₋₆ hydrocarbylgroup optionally substituted by one or more substituents selected fromhydroxy, oxo, halogen, carboxy, amino, mono- or di-C₁₋₄hydrocarbylamino, a carbocyclic or heterocyclic group with 3-6 ringmembers and containing up to 2 heteroatom ring members selected from O,N and S; and wherein one or two carbon atoms of the C₁₋₆ hydrocarbylgroup may optionally be replaced by O, S, SO, SO₂ or NH.

In one embodiment, preferred R^(10a) substituent groups on R³ (e.g. (i)when R³ is an aryl or heteroaryl group or (ii) when R³ is a non-aromaticgroup) include halogen, a group R^(a)-R^(b) wherein R^(a) is a bond, O,CO, C(X²)X¹, and R^(b) is selected from hydrogen, heterocyclic groupshaving 3-7 ring members and a C₁₋₄ hydrocarbyl group optionallysubstituted by one or more substituents selected from hydroxy, carboxy,amino, mono- or di-C₁₋₄ hydrocarbylamino, and heterocyclic groups having3-7 ring members.

Particularly preferred substituent groups R^(10a) on R³ (e.g. (i) whenR³ is an aryl or heteroaryl group or (ii) when R³ is a non-aromaticgroup) include halogen, especially fluorine, C₁₋₃ alkoxy such asmethoxy, and C₁₋₃ hydrocarbyl optionally substituted by fluorine,hydroxy (e.g. hydroxymethyl), C₁₋₂ alkoxy or a 5- or 6-memberedsaturated heterocyclic ring such as piperidino, morpholino, piperazinoand N-methylpiperazino.

In another embodiment, the substituents for R³ (whether aromatic ornon-aromatic) are selected from:

-   -   halogen (e.g. fluorine and chlorine)    -   C₁₋₄ alkoxy (e.g. methoxy and ethoxy) optionally substituted by        one or substituents selected from halogen, hydroxy, C₁₋₂ alkoxy        and five and six membered saturated heterocyclic rings        containing 1 or 2 heteroatoms selected from O, N and S, the        heterocyclic rings being optionally further substituted by one        or more C₁₋₄ groups (e.g. methyl) and wherein the S, when        present, may be present as S, SO or SO₂;    -   C₁₋₄ alkyl optionally substituted by one or substituents        selected from halogen, hydroxy, C₁₋₄ alkoxy, amino, C₁₋₄        alkylsulphonylamino, 3 to 6 membered cycloalkyl groups (e.g.        cyclopropyl), phenyl (optionally substituted by one or more        substituents selected from halogen, methyl, methoxy and amino)        and five and six membered saturated heterocyclic rings        containing 1 or 2 heteroatoms selected from O, N and S, the        heterocyclic rings being optionally further substituted by one        or more C₁₋₄ groups (e.g. methyl) and wherein the S, when        present, may be present as S, SO or SO₂;    -   hydroxy;    -   amino, mono-C₁₋₄ alkylamino, di-C₁₋₄ alkylamino,        benzyloxycarbonylamino and C₁₋₄ alkoxycarbonylamino;    -   carboxy and C₁₋₄ alkoxycarbonyl;    -   C₁₋₄ alkylaminosulphonyl and C₁₋₄ alkylsulphonylamino;    -   C₁₋₄ alkylsulphonyl;    -   a group O-Het^(s) or NH-Het^(s) where Het^(s) is a five or six        membered saturated heterocyclic ring containing 1 or 2        heteroatoms selected from O, N and S, the heterocyclic rings        being optionally further substituted by one or more C₁₋₄ groups        (e.g. methyl) and wherein the S, when present, may be present as        S, SO or SO₂;    -   five and six membered saturated heterocyclic rings containing 1        or 2 heteroatoms selected from O, N and S, the heterocyclic        rings being optionally further substituted by one or more C₁₋₄        groups (e.g. methyl) and wherein the S, when present, may be        present as S, SO or SO₂;    -   oxo; and    -   six membered aryl and heteroaryl rings containing up to two        nitrogen ring members and being optionally substituted by one or        substituents selected from halogen, methyl and methoxy.

In one preferred sub-group of compounds, R³ is a carbocyclic orheterocyclic group R^(3a) selected from phenyl; C₃₋₆ cycloalkyl; fiveand six membered saturated non-aromatic heterocyclic rings containing upto two heteroatom ring members selected from N, O, S and SO₂; sixmembered heteroaryl rings containing one, two or three nitrogen ringmembers; and five membered heteroaryl rings having up to threeheteroatom ring members selected from N, O and S;

wherein each carbocyclic or heterocyclic group R^(3a) is optionallysubstituted by up to four, preferably up to three, and more preferablyup to two (e.g. one) substituents selected from amino; hydroxy; oxo;fluorine; chlorine; C₁₋₄ alkyl-(O)_(q)— wherein q is 0 or 1 and the C₁₋₄alkyl moiety is optionally substituted by fluorine, hydroxy or C₁₋₂alkoxy; mono-C₁₋₄ alkylamino; di-C₁₋₄ alkylamino; C₁₋₄ alkoxycarbonyl;carboxy; a group R^(e)-R¹⁶ where R^(e) is a bond or a C₁₋₃ alkylenechain and R¹⁶ is selected from C₁₋₄ alkylsulphonyl; C₁₋₄alkylaminosulphonyl; C₁₋₄ alkylsulphonylamino-; amino; mono-C₁₋₄alkylamino; di-C₁₋₄ alkylamino; C₁₋₇-hydrocarbyloxycarbonylamino; sixmembered aromatic groups containing up to three nitrogen ring members;C₃₋₆ cycloalkyl; five or six membered saturated non-aromaticheterocyclic groups containing one or two heteroatom ring membersselected from N, O, S and SO₂, the group R¹⁶ when a saturatednon-aromatic group being optionally substituted by one or more methylgroups, and the group R¹⁶ when aromatic being optionally substituted byone or more groups selected from fluorine, chlorine, hydroxy, C₁₋₂alkoxy and C₁₋₂ alkyl.

In a further embodiment, R³ is selected from:

-   -   monocyclic aryl groups optionally substituted by 1-4 (for        example 1-2, e.g. 1) substituents R¹⁰ or R^(10a);    -   C₃-C₇ cycloalkyl groups optionally substituted by 1-4 (for        example 1-2, e.g. 1) substituents R¹⁰ or R^(10a);    -   saturated five membered heterocyclic rings containing 1 ring        heteroatom selected from O, N and S and being optionally        substituted by an oxo group and/or by 1-4 (for example 1-2,        e.g. 1) substituents R¹⁰ or R^(10a);    -   saturated six membered heterocyclic rings containing 1 or 2 ring        heteroatoms selected from O, N and S and being optionally        substituted by an oxo group and/or by 1-4 (for example 1-2,        e.g. 1) substituents R¹⁰ or R^(10a);    -   five membered heteroaryl rings containing 1 or 2 ring        heteroatoms selected from O, N and S and being optionally        substituted by 1-4 (for example 1-2, e.g. 1) substituents R¹⁰ or        R^(10a);    -   six membered heteroaryl rings containing 1 or 2 nitrogen ring        members (preferably 1 nitrogen ring member) and being optionally        substituted by 1-4 (for example 1-2, e.g. 1) substituents R¹⁰ or        R^(10a);    -   mono-azabicycloalkyl and diazabicycloalkyl groups each having 7        to 9 ring members and being optionally substituted by 1-4 (for        example 1-2, e.g. 1) substituents R¹⁰ or R^(10a).

Specific examples of the group Y—R³ are set out in Table 2. In Table 2,the point of attachment of the group to the nitrogen atom of thepyrazole-3-carboxamide group is represented by the terminal single bondextending from the group. Thus, by way of illustration, group CA in thetable is the 4-fluorophenyl, group CB in the table is the4-methoxybenzyl group and group CC in the table is the4-(4-methylpiperazino)-phenylmethyl group.

TABLE 2 Examples of the Group Y-R³

  CA

  CB

  CC

  CD

  CE

  CF

  CG H CH

  CI

  CJ

  CK

  CL

  CM

  CN

  CO

  CP

  CQ

  CR

  CS

  CT

  CU

  CV

  CW

  CX

  CY

  CZ

  DA

  DB

  DC

  DD

  DE

  DF

  DG

  DH

  DI

  DJ

  DK

  DL

  DM

  DN

  DO

  DP

  DQ

  DR

  DS

  DT

  DU

  DV

  DW

  DX

  DY

  DZ

  EA

  EB

  EC

  ED

  EE

  EF

  EG

  EH

  EI

  EJ

  EK

  EL

  EM

  EN

  EO

  EP

  EQ

  ER

  ES

  ET

  EU

  EV

  EW

  EX

  EY

  EZ

  FA

  FB

  FC

  FD

  FE

  FF

  FG

  FH

  FI

  FJ

  FK

  FL

  FM

  FN

One sub-set of groups selected from table 2 consists of groups CA to EU.

Another sub-set of groups selected from table 2 consists of groups CA toCV.

Preferred groups selected from Table 2 include groups CL, CM, ES, ET,FC, FG and FH.

Particularly preferred groups selected from Table 2 include groups CL,CM and ES, and most preferably CL and CM.

In another general embodiment, when R³ is an aza-cycloalkyl group, thegroup X in the compound of the formula (I) is preferably R¹-A-NR⁴wherein A is CO, NR^(g)(C═O) or O(C═O). Additionally, or alternatively,when R³ is an aza-cycloalkyl group, the nitrogen atom of theaza-cycloalkyl group is preferably not substituted with an alkylenechain linked to a 2,3-dihydro-benzo[1,4]dioxine or tetrahydronaphthalenegroup.

In another general embodiment, when Y is an alkylene chain of 1 carbonatom in length, R³ is other than an optionally substituted phenyl groupbearing a substituted or unsubstituted cyclohexyloxy or cyclohexylthiogroup.

In another general embodiment, R³ is other than a moiety containing afive membered heteroaryl ring linked directly by a single bond to amonocyclic or bicyclic aryl group or R³ is other than a moietycontaining a bis heteroaryl group comprising two five memberedheteroaryl rings linked together by a single bond.

In a further general embodiment, R¹ is other than a moiety containing afive membered heteroaryl ring linked directly by a single bond to amonocyclic or bicyclic aryl group or R¹ is other than a moietycontaining a bis heteroaryl group comprising two five memberedheteroaryl rings linked together by a single bond.

In another general embodiment, R¹-A-NR⁴ is other than an optionallysubstituted nicotinoyl-amino or benzoyl-amino group when Y—R³ is analkyl, cycloalkyl, optionally substituted phenyl or optionallysubstituted phenylalkyl group.

When A is a bond (and optionally when A is CO, NR^(g)(C═O) or O(C═O)),Y—R³ may be other than a cycloalkyl group substituted at the 1-positionwith a hydrocarbon chain simultaneously bearing an oxy substituent suchas hydroxy, an aryl substituent and a diazole or triazole substituent.

Preferably, R¹ or R³ each are other than a moiety containing asubstituted phenyl group having thio and/or oxy substituents such ashydroxy, alkoxy and alkylthio at both the 3- and 4-positions of thephenyl ring.

In a further general embodiment, when Y—R³ is unsubstituted orsubstituted benzyl or phenethyl or naphthylmethyl, X may be other thanC₁₋₅ alkylamino or C₁₋₇ acylamino.

The group Y—R³ preferably does not include a benzo-fused lactam grouphaving attached thereto an unsubstituted or substituted imidazole group.

The group Y—R³ preferably does not include the moiety —CH═C(CO₂R^(q))—S—where R^(q) is hydrogen or alkyl.

In another general embodiment, neither R¹ nor R³ contain a moiety inwhich a five membered nitrogen-containing heteroaryl group is linkeddirectly or via an alkylene, oxa-alkylene, thia-alkylene or aza-alkylenegroup to an unsubstituted pyridyl group or to a substituted aryl,heteroaryl or piperidine ring, each said ring having attached thereto asubstitutent selected from cyano, and substituted or unsubstitutedamino, aminoalkyl, amidine, guanidine, and carbamoyl groups.

In a further general embodiment, R¹ and R³ are each other than anunsaturated nitrogen-containing heterocyclic group or anitrogen-containing heteroaryl group, or a benzfuran or benzthiophenegroup wherein the said nitrogen-containing heterocyclic group,nitrogen-containing heteroaryl group, bicyclic benzfuran orbenzthiophene group are linked directly by a single bond to asubstituted pyridyl or phenyl group.

In another general embodiment, neither R¹ nor R³ contain a moiety inwhich a five membered nitrogen-containing heteroaryl group is linkeddirectly or via an alkylene, oxa-alkylene, thia-alkylene or aza-alkylenegroup to a substituted aryl, heteroaryl or piperidine group or to anunsubstituted pyridyl group.

In general, it is preferred that the compounds of the invention, wherethey contain a carboxylic acid group, contain no more than one suchgroup.

Particular and Preferred Sub-Groups of the formulae (I), (Ia) and (Ib)

One particular group of compounds of the invention is represented by theformula (II):

or salts or tautomers or N-oxides or solvates thereof;wherein R¹, R², R³ and Y are each independently selected from R¹, R², R³and Y as defined herein.

Within formula (II), it is preferred that R² is hydrogen or C₁₋₄ alkyl(e.g. C₁₋₃ alkyl), and more preferably R² is hydrogen.

In one sub-group of compounds of the formula (II), R¹ is:

(i) phenyl optionally substituted by one or more substituents (e.g. 1, 2or 3) selected from fluorine; chlorine; hydroxy; 5- and 6-memberedsaturated heterocyclic groups containing 1 or 2 heteroatoms selectedfrom O, N and S, the heterocyclic groups being optionally substituted byone or more C₁₋₄ alkyl groups; hydrocarbyloxy; and C₁₋₄ hydrocarbyl;wherein the C₁₋₄ hydrocarbyl and C₁₋₄ hydrocarbyloxy groups areoptionally substituted by one or more substituents chosen from hydroxy,fluorine, C₁₋₂ alkoxy, amino, mono and di-C₁₋₄ alkylamino, phenyl,halophenyl, saturated carbocyclic groups having 3 to 7 ring members(more preferably 4, 5 or 6 ring members, e.g. 5 or 6 ring members) orsaturated heterocyclic groups of 5 or 6 ring members and containing upto 2 heteroatoms selected from O, S and N; or2,3-dihydro-benzo[1,4]dioxine; or(ii) a monocyclic heteroaryl group containing one or two heteroatomsselected from O, S and N; or a bicyclic heteroaryl group containing asingle heteroatom selected from O, S and N; the monocyclic and bicyclicheteroaryl groups each being optionally substituted by one or moresubstituents selected from fluorine; chlorine; C₁₋₃ hydrocarbyloxy; andC₁₋₃ hydrocarbyl optionally substituted by hydroxy, fluorine, methoxy ora five or six membered saturated carbocyclic or heterocyclic groupcontaining up to two heteroatoms selected from O, S and N; or(iii) a substituted or unsubstituted cycloalkyl group having from 3 to 6ring members; or(iv) a C₁₋₄ hydrocarbyl group optionally substituted by one or moresubstituents selected from fluorine; hydroxy; C₁₋₄ hydrocarbyloxy;amino; mono- or di-C₁₋₄ hydrocarbylamino; and carbocyclic orheterocyclic groups having from 3 to 12 ring members, and wherein one ofthe carbon atoms of the hydrocarbyl group may optionally be replaced byan atom or group selected from O, NH, SO and SO₂.

Within group (i), a sub-group of groups R¹ consists of phenyl optionallysubstituted by one or more substituents selected from fluorine;chlorine; hydroxy; C₁₋₃ hydrocarbyloxy; and C₁₋₃ hydrocarbyl wherein theC₁₋₃ hydrocarbyl group is optionally substituted by one or moresubstituents chosen from hydroxy, fluorine, C₁₋₂ alkoxy, amino, mono anddi-C₁₋₄ alkylamino, saturated carbocyclic groups having 3 to 7 ringmembers (more preferably 4, 5 or 6 ring members, e.g. 5 or 6 ringmembers) or saturated heterocyclic groups of 5 or 6 ring members andcontaining up to 2 heteroatoms selected from O, S and N.

In another sub-group of compounds of the formula (II), R¹ is selectedfrom (i) and (iii) above and additionally from a sub-set (aii) wheresub-set (aii) consists of 2-furanyl, 3-furanyl, imidazolyl, 2-pyridyl,indolyl, 2-thienyl and 3-thienyl, each optionally substituted by one ormore substituents selected from fluorine, chlorine, C₁₋₃ hydrocarbyloxy,and C₁₋₃ hydrocarbyl optionally substituted by hydroxy, fluorine ormethoxy.

Within the group of compounds defined by the formula (II), where R¹ is(i) an optionally substituted phenyl group, it may be, for example, anunsubstituted phenyl group or a 2-monosubstituted, 3-monosubstituted,2,3 disubstituted, 2,5 disubstituted or 2,6 disubstituted phenyl groupor 2,3-dihydro-benzo[1,4]dioxine, where the substituents are selectedfrom halogen; hydroxyl; C₁₋₃ alkoxy; and C₁₋₃ alkyl groups wherein theC₁₋₃ alkyl group is optionally substituted by hydroxy, fluorine, C₁₋₂alkoxy, amino, mono and di-C₁₋₄ alkylamino, or saturated carbocyclicgroups having 3 to 6 ring members and/or saturated heterocyclic groupsof 5 or 6 ring members and containing 1 or 2 heteroatoms selected from Nand O.

In one embodiment, R¹ is selected from unsubstituted phenyl,2-fluorophenyl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-methylphenyl,2-(2-(pyrrolidin-1-yl)ethoxy)-phenyl, 3-fluorophenyl, 3-methoxyphenyl,2,6-difluorophenyl, 2-fluoro-6-hydroxyphenyl, 2-fluoro-3-methoxyphenyl,2-fluoro-5-methoxyphenyl, 2-chloro-6-methoxyphenyl,2-fluoro-6-methoxyphenyl, 2,6-dichlorophenyl and2-chloro-6-fluorophenyl, and is optionally further selected from5-fluoro-2-methoxyphenyl.

In another embodiment, R¹ is selected from unsubstituted phenyl,2-fluorophenyl, 2-hydroxyphenyl, 2-methoxyphenyl, 2-methylphenyl,2-(2-(pyrrolidin-1-yl)ethoxy)-phenyl, 3-fluorophenyl, 3-methoxyphenyl,2,6-difluorophenyl, 2-fluoro-6-hydroxyphenyl, 2-fluoro-3-methoxyphenyland 2-fluoro-5-methoxyphenyl.

Particular groups R¹ are 2,6-difluorophenyl, 2-fluoro-6-methoxyphenyland 2,6-dichlorophenyl.

One particularly preferred group R¹ is 2,6-difluorophenyl.

Another particularly preferred group R¹ is 2,6-dichlorophenyl.

When R¹ is (ii) a monocyclic heteroaryl group containing one or twoheteroatoms selected from O, S and N or a bicyclic heteroaryl groupcontaining a single heteroatom, examples of monocyclic and bicyclicheteroaryl groups include furanyl (e.g. 2-furanyl and 3-furanyl),imidazolyl, pyridyl (e.g. 2-pyridyl), indolyl, thienyl (e.g. 2-thienyland 3-thienyl) groups. The optional substituents for such groups caninclude chlorine, fluorine, methyl, methoxy, hydroxymethyl,methoxymethyl, morpholinomethyl, piperazinomethyl,N-methylypiperazinomethyl and piperidinylmethyl groups. Particularexamples of groups (ii) include unsubstituted 2-furanyl,3-methyl-2-furanyl, unsubstituted 4-(1H)-imidazolyl, unsubstituted5-(1H)-imidazolyl, unsubstituted 3-furanyl, unsubstituted 3-thienyl,2-methyl-3-thienyl and unsubstituted 3-pyrrolyl, and further examplesinclude 4-methoxy-3-thienyl, 5-(1-pyrrolidinyl)methyl-2-furyl and5-(4-morpholino)methyl-2-furyl groups.

When R¹ is (iii) an optionally substituted cycloalkyl group, it can befor example a substituted or unsubstituted cyclopropyl, cyclobutyl,cyclopentyl or cyclohexyl group. When the cycloalkyl group issubstituted, preferred substituents include methyl, fluorine andhydroxyl. Particular examples of cycloalkyl groups include1-methylcyclopropyl, 1-hydroxycyclopropyl, and unsubstituted cyclohexyl,cyclopentyl and cyclobutyl.

In the context of formula (II) and the group R¹, examples of optionallysubstituted hydrocarbyl groups are optionally substituted methyl, ethyland propyl groups wherein one of the carbon atoms of the hydrocarbylgroup is optionally replaced by O, NH, SO or SO₂. Particular examples ofsuch groups include methyl, ethyl, trifluoromethyl, methyl and ethylsubstituted with a carbocyclic or heterocyclic group having from 3 to 12ring members, sulphonylmethyl substituted with a carbocyclic orheterocyclic group having from 3 to 12 ring members, hydroxymethyl,hydroxyethyl, 3-hydroxy-2-propyl, propyl, isopropyl, butyl and tertiarybutyl. Examples of hydrocarbyl groups and carbocylic and heteroacyclicgroups are as set out above in the general definitions of such groups.Particular carbocyclic and heterocyclic groups include unsubstituted orsubstituted phenyl, indolyl, tetrazolyl, triazolyl, piperidinyl,morpholinyl, piperazinyl, N-methylpiperazinyl, imidazolyl wherein theoptional substituents may be selected from the group R¹⁰, and sub-groupsthereof, as defined herein.

In another sub-group of compounds of the formula (II), R¹ is a C₁₋₄hydrocarbyl group optionally substituted by one or more substituentsselected from fluorine, hydroxy, hydrocarbyloxy, amino, mono- or di-C₁₋₄hydrocarbylamino, and carbocyclic or heterocyclic groups having from 3to 12 ring members, and wherein 1 of the carbon atoms of the hydrocarbylgroup may optionally be replaced by an atom or group selected from O,NH, SO and SO₂.

In one embodiment, R¹ is a group R^(1a)—(V)_(n)— where:

n is 0 or 1;V is selected from CH₂, CH₂CH₂ and SO₂CH₂; andR^(1a) is a carbocyclic or heterocyclic group selected from phenyl; fivemembered heteroaryl rings having up to 4 heteroatom ring membersselected from N, O and S;six membered heteroaryl rings containing one or two nitrogen ringmembers; five or six membered saturated non-aromatic heterocyclic ringscontaining one or two heteroatom ring members selected from N, O, S andSO₂;C₃₋₆ cycloalkyl groups; indole; and quinoline;wherein each of the carbocyclic and heterocyclic groups R^(1a) can beoptionally substituted by one or more substituents selected from five orsix membered saturated non-aromatic carbocyclic and heterocyclic groupscontaining up to two heteroatom ring members selected from N, O, S andSO₂; hydroxy; amino; oxo; mono-C₁₋₄ alkylamino; di-C₁₋₄ alkylamino;fluorine; chlorine; nitro; C₁₋₄ alkyl-(O)_(q)— wherein q is 0 or 1 andthe C₁₋₄ alkyl moiety is optionally substituted by fluorine, hydroxy,C₁₋₂ alkoxy or a five or six membered saturated non-aromatic carbocyclicor heterocyclic group containing up to two heteroatom ring membersselected from N, O, S and SO₂; phenyl and C₁₋₂-alkylene dioxy.

Specific examples of groups R¹—CO— in formula (II) are set out in Table1 above.

One sub-group of preferred groups R¹—CO consists of the groups J, AB,AH, AJ, AL, AS, AX, AY, AZ, BA, BB, BD, BH, BL, BQ and BS.

Another sub-group of groups R¹—CO consists of the groups A to BF.

A further sub-group of groups R¹—CO consists of the groups A to BS.

Particularly preferred groups are the groups AJ, BQ and BS in Table 1,e.g. the sub-set consisting of AJ and BQ.

Another group of compounds of the invention is represented by theformula (III):

or salts or tautomers or N-oxides or solvates thereof;wherein R¹, R², R³ and Y are as defined herein.

Examples of, and preferences, for the groups R¹, R², R³ and Y are as setout above for compounds of the formulae (0), (I⁰), (I), (Ia), (Ib) and(II) unless the context indicates otherwise.

Particular sub-groups of compounds of the formula (III) include:

(i) compounds wherein R¹ is a heteroaryl group containing 1, 2 or 3heteroatom ring members selected from N, O and S;(ii) compounds wherein R¹ is a C₁₋₆ hydrocarbyl group optionallysubstituted by one or more substituents selected from fluorine, hydroxy,C₁₋₄ hydrocarbyloxy, amino, mono- or di-C₁₋₄ hydrocarbylamino, andcarbocyclic or heterocyclic groups having from 3 to 12 ring members, andwherein 1 of the carbon atoms of the hydrocarbyl group may optionally bereplaced by an atom or group selected from O, NH, SO and SO₂; and(iii) compounds wherein R¹ is a non-aromatic carbocyclic or heterocyclicgroup having from 3 to 12 ring members.

Examples of compounds of the formula (III) wherein R¹ is (i) aheteroaryl group include 5- and 6-membered monocyclic heteroaryl groups,e.g. containing for 2 heteratom ring members selected from O, N and S.In one embodiment, the heteroaryl group is a monocyclic group containing1 or 2 nitrogen ring members. In another embodiment, the heteroarylgroups are selected from 6-membered rings containing 1 or 2 nitrogenring members, for example pyridine, pyrimidine, pyrazine and pridazinegroups, one particular sub-group consisting of pyrazinyl and pyridyl.

The heteroaryl groups can be unsubstituted or substituted by one or moregroups R¹⁰ as defined herein.

Examples of compounds of the formula (III) wherein R¹ is (ii) anoptionally substituted C₁₋₆ hydrocarbyl group include those in which thehydrocarbyl group is unsubstituted hydrocarbyl, for exampleunsubstituted alkyl such as methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, 1-pentyl, 2-pentyl and 3-pentyl.

Examples of compounds wherein R¹ is a non-aromatic carbocyclic orheterocyclic group include those wherein the carbocyclic or heterocylicgroup is monocyclic and contains up to 2 heteroatoms selected fromoxygen and nitrogen. Particular examples of such groups are cyclohexyland piperidino.

Another sub-group of compounds of the formula (I) can be represented bythe formula (IV):

or salts or tautomers or N-oxides or solvates thereof;wherein R¹ and R² are as defined herein;an optional second bond may be present between carbon atoms numbered 1and 2; one of U and T is selected from CH₂, CHR¹³, CR¹¹R¹³, NR¹⁴,N(O)R¹⁵, O and S(O)_(t); and the other of U and T is selected from,NR¹⁴, O, CH₂, CHR¹¹, C(R¹¹)₂, and C═O; r is 0, 1, 2, 3 or 4; t is 0, 1or 2;R¹¹ is selected from hydrogen, halogen (particularly fluorine), C₁₋₃alkyl (e.g. methyl) and C₁₋₃ alkoxy (e.g. methoxy);R¹³ is selected from hydrogen, NHR¹⁴, NOH, NOR¹⁴ and R^(a)-R^(b);R¹⁴ is selected from hydrogen and R^(d)-R^(b);R^(d) is selected from a bond, CO, C(X²)X¹, SO₂ and SO₂NR^(c);R^(a), R^(b) and R^(c) are as hereinbefore defined; andR¹⁵ is selected from C₁₋₄ saturated hydrocarbyl optionally substitutedby hydroxy, C₁₋₂ alkoxy, halogen or a monocyclic 5- or 6-memberedcarbocyclic or heterocyclic group, provided that U and T cannot be Osimultaneously.

Examples of, and preferences, for the groups R¹ and R² are as set outabove for compounds of the formulae (I), (Ia), (Ib) and (II) unless thecontext indicates otherwise.

Within formula (IV), r can be 0, 1, 2, 3 or 4. In one embodiment, r is0. In another embodiment, r is 2, and in a further embodiment r is 4.

Within formula (IV), one sub-set of preferred compounds is the set ofcompounds where there is only a single bond between the carbon atomsnumbered 1 and 2.

However, in another sub-set of compounds, there is a double bond betweenthe carbon atoms numbered 1 and 2.

Another sub-set of compounds is characterised by gem disubstitution atthe 2-carbon (when there is a single bond between carbon atoms numbers 1and 2) and/or the 6-carbon. Preferred gem disubstituents includedifluoro and dimethyl.

A further sub-set of compounds is characterised by the presence of analkoxy group, for example a methoxy group at the carbon atom numbered 3,i.e. at a position α with respect to the group T.

Within formula (IV) are compounds wherein, for example, R³ is selectedfrom any of the following ring systems:

Preferred ring systems include G1 and G3.

A preferred sub-group of compounds within formula (IV) can berepresented by the formula (IVa):

or salts or tautomers or N-oxides or solvates thereof;wherein R¹ and R² are as hereinbefore defined;one of U and T is selected from CH₂, CHR¹³, CR¹¹R¹³, NR¹⁴, N(O)R¹⁵, Oand S(O)_(t); and the other of U and T is selected from CH₂, CHR¹¹,C(R¹¹)₂, and C═O; r is 0, 1 or 2; t is 0, 1 or 2;R¹¹ is selected from hydrogen and C₁₋₃ alkyl;R¹³ is selected from hydrogen and R^(a)-R^(b);R¹⁴ is selected from hydrogen and R^(d)-R^(b);R^(d) is selected from a bond, CO, C(X²)X¹, SO₂ and SO₂NR^(c);R^(a), R^(b) and R^(c) are as hereinbefore defined; andR¹⁵ is selected from C₁₋₄ saturated hydrocarbyl optionally substitutedby hydroxy,

C₁₋₂ alkoxy, halogen or a monocyclic 5- or 6-membered carbocyclic orheterocyclic group.

Examples of, and preferences, for the groups R¹ and R² are as set outabove for compounds of the formulae (0), (I⁰), (I), (Ia), (Ib) and (II)unless the context indicates otherwise.

In formula (IVa), T is preferably selected from CH₂, CHR¹³, CR¹¹R¹³,NR¹⁴, N(O)R¹⁵, O and S(O)_(t); and U is preferably selected from CH₂,CHR¹¹, C(R¹¹)₂, and C═O.

In the definitions for substituents R¹¹ and R¹⁴, R^(b) is preferablyselected from hydrogen; monocyclic carbocyclic and heterocyclic groupshaving from 3 to 7 ring members; and C₁₋₄ hydrocarbyl (more preferablyacyclic saturated C₁₋₄ groups) optionally substituted by one or moresubstituents selected from hydroxy, oxo, halogen, amino, mono- ordi-C₁₋₄ hydrocarbylamino, and monocyclic carbocyclic and heterocyclicgroups having from 3 to 7 ring members (more preferably 3 to 6 ringmembers) and wherein one or more carbon atoms of the C₁₋₄ hydrocarbylgroup may optionally be replaced by O, S, SO, SO₂, NR^(c), X¹C(X²),C(X²)X¹;

R^(c) is selected from hydrogen and C₁₋₄ hydrocarbyl; and

-   -   X¹ is O, S or NR^(c) and X² is ═O, ═S or ═NR^(c).

R¹¹ is preferably selected from hydrogen and methyl and most preferablyis hydrogen.

R¹³ is preferably selected from hydrogen; hydroxy; halogen; cyano;amino; mono-C₁₋₄ saturated hydrocarbylamino; di-C₁₋₄ saturatedhydrocarbylamino; monocyclic 5- or 6-membered carbocyclic andheterocyclic groups; C₁₋₄ saturated hydrocarbyl optionally substitutedby hydroxy, C₁₋₂ alkoxy, halogen or a monocyclic 5- or 6-memberedcarbocyclic or heterocyclic group.

Particular examples of R¹³ are hydrogen, hydroxy, amino, C₁₋₂ alkylamino(e.g. methylamino) C₁₋₄ alkyl (e.g. methyl, ethyl, propyl and butyl),C₁₋₂ alkoxy (e.g. methoxy), C₁₋₂ alkylsulphonamido (e.g.methanesulphonamido), hydroxy-C₁₋₂ alkyl (e.g. hydroxymethyl),C₁₋₂-alkoxy-C₁₋₂ alkyl (e.g. methoxymethyl and methoxyethyl), carboxy,C₁₋₄ alkoxycarbonyl (e.g. ethoxycarbonyl) and amino-C₁₋₂-alkyl (e.g.aminomethyl).

Particular examples of R¹⁴ are hydrogen; C₁₋₄ alkyl optionallysubstituted by fluoro or a five or six membered saturated heterocyclicgroup (e.g. a group selected from (i) methyl, ethyl, n-propyl, i-propyl,butyl, 2,2,2-trifluoroethyl and tetrahydrofuranylmethyl; and/or (ii)2-fluoroethyl and 2,2-difluoroethyl); cyclopropylmethyl; substituted orunsubstituted pyridyl-C₁₋₂ alkyl (e.g. 2-pyridylmethyl); substituted orunsubstituted phenyl-C₁₋₂ alkyl (e.g. benzyl); C₁₋₄ alkoxycarbonyl (e.g.ethoxycarbonyl and t-butyloxycarbonyl); substituted and unsubstitutedphenyl-C₁₋₂ alkoxycarbonyl (e.g. benzyloxycarbonyl); substituted andunsubstituted 5- and 6-membered heteroaryl groups such as pyridyl (e.g.2-pyridyl and 6-chloro-2-pyridyl) and pyrimidinyl (e.g. 2-pyrimidinyl);C₁₋₂-alkoxy-C₁₋₂ alkyl (e.g. methoxymethyl and methoxyethyl); C₁₋₄alkylsulphonyl (e.g. methanesulphonyl).

Preferred compounds include those in which (i) U is CHR¹³ (morepreferably CH₂) and T is NR₁₄, and (ii) T is CHR¹³ (more preferably CH₂)and U is NR¹⁴.

One particular preferred sub-group of compounds of the formula (IV) canbe represented by the formula (Va):

or salts or tautomers or N-oxides or solvates thereof;wherein R^(14a) is selected from hydrogen, C₁₋₄ alkyl optionallysubstituted by fluoro (e.g. methyl, ethyl, n-propyl, i-propyl, butyl and2,2,2-trifluoroethyl), cyclopropylmethyl, phenyl-C₁₋₂ alkyl (e.g.benzyl), C₁₋₄ alkoxycarbonyl (e.g. ethoxycarbonyl andt-butyloxycarbonyl), phenyl-C₁₋₂ alkoxycarbonyl (e.g.benzyloxycarbonyl), C₁₋₂-alkoxy-C₁₋₂ alkyl (e.g. methoxymethyl andmethoxyethyl), and C₁₋₄ alkylsulphonyl (e.g. methanesulphonyl), whereinthe phenyl moieties when present are optionally substituted by one tothree substituents selected from fluorine, chlorine, C₁₋₄ alkoxyoptionally substituted by fluoro or C₁₋₂-alkoxy, and C₁₋₄ alkyloptionally substituted by fluoro or C₁₋₂-alkoxy;w is 0, 1, 2 or 3;R² is hydrogen or methyl, most preferably hydrogen;R¹¹ and r are as hereinbefore defined; andR¹⁹ is selected from fluorine; chlorine; C₁₋₄ alkoxy optionallysubstituted by fluoro or C₁₋₂-alkoxy; and C₁₋₄ alkyl optionallysubstituted by fluoro or C₁₋₂-alkoxy.

Another particular preferred sub-group of compounds of the formula (IV)can be represented by the formula (Vb):

or salts or tautomers or N-oxides or solvates thereof;wherein R^(14a) is selected from hydrogen, C₁₋₄ alkyl optionallysubstituted by fluoro (e.g. methyl, ethyl, n-propyl, i-propyl, butyl and2,2,2-trifluoroethyl), cyclopropylmethyl, phenyl-C₁₋₂ alkyl (e.g.benzyl), C₁₋₄ alkoxycarbonyl (e.g. ethoxycarbonyl andt-butyloxycarbonyl), phenyl-C₁₋₂ alkoxycarbonyl (e.g.benzyloxycarbonyl), C₁₋₂-alkoxy-C₁₋₂ alkyl (e.g. methoxymethyl andmethoxyethyl), and C₁₋₄ alkylsulphonyl (e.g. methanesulphonyl), whereinthe phenyl moieties when present are optionally substituted by one tothree substituents selected from fluorine, chlorine, C₁₋₄ alkoxyoptionally substituted by fluoro or C₁₋₂-alkoxy, and C₁₋₄ alkyloptionally substituted by fluoro or C₁₋₂-alkoxy;w is 0, 1, 2 or 3;R² is hydrogen or methyl, most preferably hydrogen;R¹¹ and r are as hereinbefore defined; andR¹⁹ is selected from fluorine; chlorine; C₁₋₄ alkoxy optionallysubstituted by fluoro or C₁₋₂-alkoxy; and C₁₋₄ alkyl optionallysubstituted by fluoro or C₁₋₂-alkoxy.

In formulae (Va) and (Vb), when w is 1, 2 or 3, it is preferred that thephenyl ring is 2-monosubstituted, 3-monosubstituted, 2,6-disubstituted,2,3-disubstituted, 2,4-disubstituted 2,5-disubstituted,2,3,6-trisubstituted or 2,4,6-trisubstituted. Most preferably the phenylring is disubstituted at positions 2- and 6- with substituents selectedfrom fluorine, chlorine and methoxy.

R¹¹ is preferably hydrogen (or r is 0).

R^(14a) is most preferably hydrogen or methyl.

One preferred sub-group of compounds of the formula (Va) can berepresented by the formula (VIa):

or salts or tautomers or N-oxides or solvates thereof;wherein R²⁰ is selected from hydrogen and methyl;R²¹ is selected from fluorine and chlorine; andR²² is selected from fluorine, chlorine and methoxy; orone of R²¹ and R²² is hydrogen and the other is selected from chlorine,methoxy, ethoxy, difluoromethoxy, trifluoromethoxy and benzyloxy.

Another preferred sub-group of compounds of the formula (Va) can berepresented by the formula (VIb):

or salts or tautomers or N-oxides or solvates thereof;wherein R²⁰ is selected from hydrogen and methyl;R^(21a) is selected from fluorine and chlorine; andR^(22a) is selected from fluorine, chlorine and methoxy.

Particular compounds within formula (VIb) include:

-   4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid    piperidin-4-ylamide;-   4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid    (1-methyl-piperidin-4-yl)-amide;-   4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid    piperidin-4-ylamide;-   and-   4-(2-fluoro-6-methoxy-benzoylamino)-1H-pyrazole-3-carboxylic acid    piperidin-4-ylamide;    or salts or tautomers or N-oxides or solvates thereof.

A further group of compounds of the invention is represented by theformula (VII):

or salts or tautomers or N-oxides or solvates thereof;wherein R², R³ and Y are as hereinbefore defined and G is a 5- or6-membered carbocyclic or heterocyclic ring.

The group G can be an unsubstituted carbocyclic or heterocyclic ring orit can be a substituted carbocyclic or heterocyclic ring bearing one ormore substituents selected from the groups R¹⁰ and R^(10a) ashereinbefore defined

The carbocyclic or heterocyclic ring may be aromatic or non-aromatic andexamples of such heterocyclic rings are set out above. In the context ofthe group G, preferred heterocyclic rings are those containing anitrogen ring atom through which the group G is connected to thepyrazole ring. Particular heterocyclic rings are saturated heterocyclicrings containing up to 3 nitrogen atoms (more usually up to 2, forexample 1) and optionally an oxygen atom. Particular examples of suchrings are six membered rings such as piperidine, piperazine, N-methylpiperazine and morpholine.

When the group G is a carbocyclic group, it can be, for example a6-membered aryl ring. For example, the group G can be an unsubstitutedphenyl group or it can be a substituted phenyl group bearing one or moresubstituents selected from the groups R¹⁰ and R^(10a) as hereinbeforedefined. The substituents, when present, are more typically smallsubstituents such as hydroxyl, halogen (e.g. fluorine and chlorine), andC₁₋₄ hydrocarbyl (methyl, ethyl and cyclopropyl) optionally substitutedby fluorine (e.g. trifluoromethyl) or hydroxy (e.g. hydroxymethyl).

In one general embodiment, when X is a non-aromatic heterocyclic group,then R³ may be other than a six membered monocyclic aryl or heteroarylgroup linked directly to a 5,6-fused bicyclic heteroaryl group.

A further group of compounds of the invention is represented by theformula (VIII):

or salts or tautomers or N-oxides or solvates thereof;wherein R¹, R², R³ and Y are as defined herein.

Preferred groups R¹, R², Y and R³ are as set out above in the sectionheaded “General Preferences and Definitions” and in relation tocompounds of the formulae (I) and (II) and sub-groups thereof as definedherein.

For the avoidance of doubt, it is to be understood that each general andspecific preference, embodiment and example of the groups R¹ may becombined with each general and specific preference, embodiment andexample of the groups R² and/or R³ and/or R⁴ and/or R¹⁰ and/or Y and/orR^(g) and/or sub-groups thereof as defined herein and that all suchcombinations are embraced by this application.

The various functional groups and substituents making up the compoundsof the formula (I) are typically chosen such that the molecular weightof the compound of the formula (I) does not exceed 1000. More usually,the molecular weight of the compound will be less than 750, for exampleless than 700, or less than 650, or less than 600, or less than 550.More preferably, the molecular weight is less than 525 and, for example,is 500 or less.

Particular compounds of the invention are as illustrated in the examplesbelow.

Salts, Solvates, Tautomers, Isomers, N-Oxides, Esters, Prodrugs andIsotopes

Unless otherwise specified, a reference to a particular compound alsoincludes ionic, salt, solvate, and protected forms thereof, for example,as discussed below.

Many compounds of the formula (I) can exist in the form of salts, forexample acid addition salts or, in certain cases salts of organic andinorganic bases such as carboxylate, sulphonate and phosphate salts. Allsuch salts are within the scope of this invention, and references tocompounds of the formula (I) include the salt forms of the compounds. Asin the preceding sections of this application, all references to formula(I) should be taken to refer also to formulae (0), (I⁰), (I), (Ia),(Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or(VIII) and sub-groups thereof unless the context indicates otherwise.

Salt forms may be selected and prepared according to methods describedin Pharmaceutical Salts Properties, Selection, and Use, P. HeinrichStahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639-026-8,Hardcover, 388 pages, August 2002.

Acid addition salts may be formed with a wide variety of acids, bothinorganic and organic. Examples of acid addition salts include saltsformed with an acid selected from the group consisting of acetic,2,2-dichloroacetic, adipic, alginic, ascorbic (e.g. L-ascorbic),L-aspartic, benzenesulphonic, benzoic, 4-acetamidobenzoic, butanoic, (+)camphoric, camphor-sulphonic, (+)-(1S)-camphor-10-sulphonic, capric,caproic, caprylic, cinnamic, citric, cyclamic, dodecylsulphuric,ethane-1,2-disulphonic, ethanesulphonic, 2-hydroxyethanesulphonic,formic, fumaric, galactaric, gentisic, glucoheptonic, D-gluconic,glucuronic (e.g. D-glucuronic), glutamic (e.g. L-glutamic),α-oxoglutaric, glycolic, hippuric, hydrobromic, hydrochloric, hydriodic,isethionic, (+)-L-lactic, (±)-DL-lactic, lactobionic, maleic, malic,(−)-L-malic, malonic, (±)-DL-mandelic, methanesulphonic,naphthalene-2-sulphonic, naphthalene-1,5-disulphonic,1-hydroxy-2-naphthoic, nicotinic, nitric, oleic, orotic, oxalic,palmitic, pamoic, phosphoric, propionic, L-pyroglutamic, salicylic,4-amino-salicylic, sebacic, stearic, succinic, sulphuric, tannic,(+)-L-tartaric, thiocyanic, p-toluenesulphonic, undecylenic and valericacids, as well as acylated amino acids and cation exchange resins.

One particular group of salts consists of salts formed fromhydrochloric, hydriodic, phosphoric, nitric, sulphuric, citric, lactic,succinic, maleic, malic, isethionic, fumaric, benzenesulphonic,toluenesulphonic, methanesulphonic, ethanesulphonic,naphthalenesulphonic, valeric, acetic, propanoic, butanoic, malonic,glucuronic and lactobionic acids.

One preferred group of salts consists of salts formed from hydrochloric,acetic, adipic, L-aspartic and DL-lactic acids.

Particularly preferred salts are hydrochloride salts

For example, if the compound is anionic, or has a functional group whichmay be anionic (e.g., —COOH may be —COO⁻), then a salt may be formedwith a suitable cation. Examples of suitable inorganic cations include,but are not limited to, alkali metal ions such as Na⁺ and K⁺, alkalineearth cations such as Ca²⁺ and Mg²⁺, and other cations such as Al³⁺.Examples of suitable organic cations include, but are not limited to,ammonium ion (i.e., NH₄ ⁺) and substituted ammonium ions (e.g., NH₃R⁺,NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺). Examples of some suitable substituted ammoniumions are those derived from: ethylamine, diethylamine,dicyclohexylamine, triethylamine, butylamine, ethylenediamine,ethanolamine, diethanolamine, piperazine, benzylamine,phenylbenzylamine, choline, meglumine, and tromethamine, as well asamino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

Where the compounds of the formula (I) contain an amine function, thesemay form quaternary ammonium salts, for example by reaction with analkylating agent according to methods well known to the skilled person.Such quaternary ammonium compounds are within the scope of formula (I).

The salt forms of the compounds of the invention are typicallypharmaceutically acceptable salts, and examples of pharmaceuticallyacceptable salts are discussed in Berge et al., 1977, “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19. However, saltsthat are not pharmaceutically acceptable may also be prepared asintermediate forms which may then be converted into pharmaceuticallyacceptable salts. Such non-pharmaceutically acceptable salts forms,which may be useful, for example, in the purification or separation ofthe compounds of the invention, also form part of the invention.

Compounds of the formula (I) containing an amine function may also formN-oxides. A reference herein to a compound of the formula (I) thatcontains an amine function also includes the N-oxide.

Where a compound contains several amine functions, one or more than onenitrogen atom may be oxidised to form an N-oxide. Particular examples ofN-oxides are the N-oxides of a tertiary amine or a nitrogen atom of anitrogen-containing heterocycle.

N-Oxides can be formed by treatment of the corresponding amine with anoxidizing agent such as hydrogen peroxide or a per-acid (e.g. aperoxycarboxylic acid), see for example Advanced Organic Chemistry, byJerry March, 4^(th) Edition, Wiley Interscience, pages. Moreparticularly, N-oxides can be made by the procedure of L. W. Deady (Syn.Comm. 1977, 7, 509-514) in which the amine compound is reacted withm-chloroperoxybenzoic acid (MCPBA), for example, in an inert solventsuch as dichloromethane.

Compounds of the formula (I) may exist in a number of differentgeometric isomeric, and tautomeric forms and references to compounds ofthe formula (I) include all such forms. For the avoidance of doubt,where a compound can exist in one of several geometric isomeric ortautomeric forms and only one is specifically described or shown, allothers are nevertheless embraced by formula (I).

For example, in compounds of the formula (I) the pyrazole group may takeeither of the following two tautomeric forms A and B. For simplicity,the general formula (I) illustrates form A but the formula is to betaken as embracing both tautomeric forms.

Other examples of tautomeric forms include, for example, keto-, enol-,and enolate-forms, as in, for example, the following tautomeric pairs:keto/enol (illustrated below), imine/enamine, amide/imino alcohol,amidine/amidine, nitroso/oxime, thioketone/enethiol, andnitro/aci-nitro.

Where compounds of the formula (I) contain one or more chiral centres,and can exist in the form of two or more optical isomers, references tocompounds of the formula (I) include all optical isomeric forms thereof(e.g. enantiomers, epimers and diastereoisomers), either as individualoptical isomers, or mixtures (e.g. racemic mixtures) or two or moreoptical isomers, unless the context requires otherwise.

The optical isomers may be characterised and identified by their opticalactivity (i.e. as + and − isomers, or d and l isomers) or they may becharacterised in terms of their absolute stereochemistry using the “Rand S” nomenclature developed by Cahn, Ingold and Prelog, see AdvancedOrganic Chemistry by Jerry March, 4^(th) Edition, John Wiley & Sons, NewYork, 1992, pages 109-114, and see also Cahn, Ingold & Prelog, Angew.Chem. Int. Ed. Engl., 1966, 5, 385-415.

Optical isomers can be separated by a number of techniques includingchiral chromatography (chromatography on a chiral support) and suchtechniques are well known to the person skilled in the art.

Where compounds of the formula (I) exist as two or more optical isomericforms, one enantiomer in a pair of enantiomers may exhibit advantagesover the other enantiomer, for example, in terms of biological activity.Thus, in certain circumstances, it may be desirable to use as atherapeutic agent only one of a pair of enantiomers, or only one of aplurality of diastereoisomers. Accordingly, the invention providescompositions containing a compound of the formula (I) having one or morechiral centres, wherein at least 55% (e.g. at least 60%, 65%, 70%, 75%,80%, 85%, 90% or 95%) of the compound of the formula (I) is present as asingle optical isomer (e.g. enantiomer or diastereoisomer). In onegeneral embodiment, 99% or more (e.g. substantially all) of the totalamount of the compound of the formula (I) may be present as a singleoptical isomer (e.g. enantiomer or diastereoisomer).

The compounds of the invention include compounds with one or moreisotopic substitutions, and a reference to a particular element includeswithin its scope all isotopes of the element. For example, a referenceto hydrogen includes within its scope ¹H, ²H (D), and ³H (T). Similarly,references to carbon and oxygen include within their scope respectively¹²C, ¹³C and ¹⁴C and ¹⁶O and ¹⁸O.

The isotopes may be radioactive or non-radioactive. In one embodiment ofthe invention, the compounds contain no radioactive isotopes. Suchcompounds are preferred for therapeutic use. In another embodiment,however, the compound may contain one or more radioisotopes. Compoundscontaining such radioisotopes may be useful in a diagnostic context.

Esters such as carboxylic acid esters and acyloxy esters of thecompounds of formula (I) bearing a carboxylic acid group or a hydroxylgroup are also embraced by Formula (I). Examples of esters are compoundscontaining the group —C(═O)OR, wherein R is an ester substituent, forexample, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀ arylgroup, preferably a C₁₋₇ alkyl group. Particular examples of estergroups include, but are not limited to, —C(═O)OCH₃, —C(═O)OCH₂CH₃,—C(═O)OC(CH₃)₃, and —C(═O)OPh. Examples of acyloxy (reverse ester)groups are represented by —OC(═O)R, wherein R is an acyloxy substituent,for example, a C₁₋₇ alkyl group, a C₃₋₂₀ heterocyclyl group, or a C₅₋₂₀aryl group, preferably a C₁₋₇ alkyl group. Particular examples ofacyloxy groups include, but are not limited to, —OC(═O)CH₃ (acetoxy),—OC(═O)CH₂CH₃, —OC(═O)C(CH₃)₃, —OC(═O)Ph, and —OC(═O)CH₂Ph.

Also encompassed by formula (I) are any polymorphic forms of thecompounds, solvates (e.g. hydrates), complexes (e.g. inclusion complexesor clathrates with compounds such as cyclodextrins, or complexes withmetals) of the compounds, and pro-drugs of the compounds. By “prodrugs”is meant for example any compound that is converted in vivo into abiologically active compound of the formula (I).

For example, some prodrugs are esters of the active compound (e.g., aphysiologically acceptable metabolically labile ester). Duringmetabolism, the ester group (—C(═O)OR) is cleaved to yield the activedrug. Such esters may be formed by esterification, for example, of anyof the carboxylic acid groups (—C(═O)OH) in the parent compound, with,where appropriate, prior protection of any other reactive groups presentin the parent compound, followed by deprotection if required.

Examples of such metabolically labile esters include those of theformula —C(═O)OR wherein R is:

C₁₋₇ alkyl(e.g., -Me, -Et, -nPr, -iPr, -nBu, -sBu, -iBu, -tBu);C₁₋₇ aminoalkyl(e.g., aminoethyl; 2-(N,N-diethylamino)ethyl; 2-(4-morpholino)ethyl);andacyloxy-C₁₋₇alkyl(e.g., acyloxymethyl;acyloxyethyl;pivaloyloxymethyl;acetoxymethyl;1-acetoxyethyl;1-(1-methoxy-1-methyl)ethyl-carbonxyloxyethyl;1-(benzoyloxy)ethyl; isopropoxy-carbonyloxymethyl;1-isopropoxy-carbonyloxyethyl; cyclohexyl-carbonyloxymethyl;1-cyclohexyl-carbonyloxyethyl;cyclohexyloxy-carbonyloxymethyl;1-cyclohexyloxy-carbonyloxyethyl;(4-tetrahydropyranyloxy)carbonyloxymethyl;1-(4-tetrahydropyranyloxy)carbonyloxyethyl;(4-tetrahydropyranyl)carbonyloxymethyl; and1-(4-tetrahydropyranyl)carbonyloxyethyl).

Also, some prodrugs are activated enzymatically to yield the activecompound, or a compound which, upon further chemical reaction, yieldsthe active compound (for example, as in ADEPT, GDEPT, LIDEPT, etc.). Forexample, the prodrug may be a sugar derivative or other glycosideconjugate, or may be an amino acid ester derivative.

Biological Activity

The compounds of the formulae (0), (I⁰), (I), (Ia), (Ib), (II), (III),(IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groupsthereof are inhibitors of cyclin dependent kinases, and in particularcyclin dependent kinases selected from CDK1, CDK2, CDK3, CDK4, CDK5 andCDK6.

Preferred compounds are compounds that inhibit one or more CDK kinasesselected from CDK1, CDK2, CDK4 and CDK5, for example CDK1 and/or CDK2.

The compounds of the invention are also considered to be inhibitors ofglycogen synthase kinase-3 (GSK3).

As a consequence of their activity in modulating or inhibiting CDKkinases and glycogen synthase kinase, they are expected to be useful inproviding a means of arresting, or recovering control of, the cell cyclein abnormally dividing cells. It is therefore anticipated that thecompounds will prove useful in treating or preventing proliferativedisorders such as cancers. It is also envisaged that the compounds ofthe invention will be useful in treating conditions such as viralinfections, type II or non-insulin dependent diabetes mellitus,autoimmune diseases, head trauma, stroke, epilepsy, neurodegenerativediseases such as Alzheimer's, motor neurone disease, progressivesupranuclear palsy, corticobasal degeneration and Pick's disease forexample. One sub-group of disease states and conditions where it isenvisaged that the compounds of the invention will be useful consists ofviral infections, autoimmune diseases and neurodegenerative diseases.

CDKs play a role in the regulation of the cell cycle, apoptosis,transcription, differentiation and CNS function. Therefore, CDKinhibitors could be useful in the treatment of diseases in which thereis a disorder of proliferation, apoptosis or differentiation such ascancer. In particular RB+ve tumours may be particularly sensitive to CDKinhibitors. RB−ve tumours may also be sensitive to CDK inhibitors.

Examples of cancers which may be inhibited include, but are not limitedto, a carcinoma, for example a carcinoma of the bladder, breast, colon(e.g. colorectal carcinomas such as colon adenocarcinoma and colonadenoma), kidney, epidermis, liver, lung, for example adenocarcinoma,small cell lung cancer and non-small cell lung carcinomas, oesophagus,gall bladder, ovary, pancreas e.g. exocrine pancreatic carcinoma,stomach, cervix, thyroid, prostate, or skin, for example squamous cellcarcinoma; a hematopoietic tumour of lymphoid lineage, for exampleleukemia, acute lymphocytic leukemia, B-cell lymphoma, T-cell lymphoma,Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy cell lymphoma, orBurkett's lymphoma; a hematopoietic tumour of myeloid lineage, forexample acute and chronic myelogenous leukemias, myelodysplasticsyndrome, or promyelocytic leukemia; thyroid follicular cancer; a tumourof mesenchymal origin, for example fibrosarcoma or habdomyosarcoma, atumour of the central or peripheral nervous system, for exampleastrocytoma, neuroblastoma, glioma or schwannoma; melanoma; seminoma;teratocarcinoma; osteosarcoma; xeroderma pigmentosum; keratoctanthoma;thyroid follicular cancer; or Kaposi's sarcoma.

The cancers may be cancers which are sensitive to inhibition of any oneor more cyclin dependent kinases selected from CDK1, CDK2, CDK3, CDK4,CDK5 and CDK6, for example, one or more CDK kinases selected from CDK1,CDK2, CDK4 and CDK5, e.g. CDK1 and/or CDK2.

Whether or not a particular cancer is one which is sensitive toinhibition by a cyclin dependent kinase may be determined by means of acell growth assay as set out in Example 250 below or by a method as setout in the section headed “Methods of Diagnosis”.

CDKs are also known to play a role in apoptosis, proliferation,differentiation and transcription and therefore CDK inhibitors couldalso be useful in the treatment of the following diseases other thancancer; viral infections, for example herpes virus, pox virus,Epstein-Barr virus, Sindbis virus, adenovirus, HIV, HPV, HCV and HCMV;prevention of AIDS development in HIV-infected individuals; chronicinflammatory diseases, for example systemic lupus erythematosus,autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis,inflammatory bowel disease, and autoimmune diabetes mellitus;cardiovascular diseases for example cardiac hypertrophy, restenosis,atherosclerosis; neurodegenerative disorders, for example Alzheimer'sdisease, AIDS-related dementia, Parkinson's disease, amyotropic lateralsclerosis, retinitis pigmentosa, spinal muscular atropy and cerebellardegeneration; glomerulonephritis; myelodysplastic syndromes, ischemicinjury associated myocardial infarctions, stroke and reperfusion injury,arrhythmia, atherosclerosis, toxin-induced or alcohol related liverdiseases, haematological diseases, for example, chronic anemia andaplastic anemia; degenerative diseases of the musculoskeletal system,for example, osteoporosis and arthritis, aspirin-sensitiverhinosinusitis, cystic fibrosis, multiple sclerosis, kidney diseases andcancer pain.

It has also been discovered that some cyclin-dependent kinase inhibitorscan be used in combination with other anticancer agents. For example,the cyclin-dependent kinase inhibitor flavopiridol has been used withother anticancer agents in combination therapy.

Thus, in the pharmaceutical compositions, uses or methods of thisinvention for treating a disease or condition comprising abnormal cellgrowth, the disease or condition comprising abnormal cell growth in oneembodiment is a cancer.

One group of cancers includes human breast cancers (e.g. primary breasttumours, node-negative breast cancer, invasive duct adenocarcinomas ofthe breast, non-endometrioid breast cancers); and mantle cell lymphomas.In addition, other cancers are colorectal and endometrial cancers.

Another sub-set of cancers includes breast cancer, ovarian cancer, coloncancer, prostate cancer, oesophageal cancer, squamous cancer andnon-small cell lung carcinomas.

The activity of the compounds of the invention as inhibitors of cyclindependent kinases and glycogen synthase kinase-3 can be measured usingthe assays set forth in the examples below and the level of activityexhibited by a given compound can be defined in terms of the IC₅₀ value.Preferred compounds of the present invention are compounds having anIC₅₀ value of less than 1 micromole, more preferably less than 0.1micromole.

Methods for the Preparation of Compounds of the Invention

Compounds of the formula (I) and the various sub-groups thereof can beprepared in accordance with synthetic methods well known to the skilledperson. Unless stated otherwise, R¹, R², R³, Y, X and A are ashereinbefore defined.

In this section, as in all the other sections of this application,references to formula (I) should be taken to refer also to formulae (0),(I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa),(VIb), (VII) or (VIII) and sub-groups thereof unless the contextindicates otherwise.

Compounds of the formula (I) wherein R¹-A- forms an acyl group R¹—CO—can be prepared by reacting a carboxylic acid of the formula R¹—CO₂H oran activated derivative thereof with an appropriately substituted4-amino-pyrazole as shown in Scheme 1.

The starting material for the synthetic route shown in Scheme 1 is the4-nitro-pyrazole-3-carboxylic acid (X) which can either be obtainedcommercially or can be prepared by nitration of the corresponding4-unsubstituted pyrazole carboxy compound.

The 4-nitro-pyrazole carboxylic acid (X), or a reactive derivativethereof, is reacted with the amine H₂N—Y—R³ to give the 4-nitro-amide(XI). The coupling reaction between the carboxylic acid (X) and theamine is preferably carried out in the presence of a reagent of the typecommonly used in the formation of peptide linkages. Examples of suchreagents include 1,3-dicyclohexylcarbodiimide (DCC) (Sheehan et al, J.Amer. Chem. Soc. 1955, 77, 1067),1-ethyl-3-(3′-dimethylaminopropyl)-carbodiimide (referred to hereineither as EDC or EDAC but also known in the art as EDCI and WSCDI)(Sheehan et al, J. Org. Chem., 1961, 26, 2525), uronium-based couplingagents such as O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) and phosphonium-based coupling agents such as1-benzo-triazolyloxytris-(pyrrolidino)phosphonium hexafluorophosphate(PyBOP) (Castro et al, Tetrahedron Letters, 1990, 31, 205).Carbodiimide-based coupling agents are advantageously used incombination with 1-hydroxy-7-azabenzotriazole (HOAt) (L. A. Carpino, J.Amer. Chem. Soc., 1993, 115, 4397) or 1-hydroxybenzotriazole (HOBt)(Konig et al, Chem. Ber., 103, 708, 2024-2034). Preferred couplingreagents include EDC (EDAC) and DCC in combination with HOAt or HOBt.

The coupling reaction is typically carried out in a non-aqueous,non-protic solvent such as acetonitrile, dioxan, dimethylsulphoxide,dichloromethane, dimethylformamide or N-methylpyrrolidine, or in anaqueous solvent optionally together with one or more miscibleco-solvents. The reaction can be carried out at room temperature or,where the reactants are less reactive (for example in the case ofelectron-poor anilines bearing electron withdrawing groups such assulphonamide groups) at an appropriately elevated temperature. Thereaction may be carried out in the presence of a non-interfering base,for example a tertiary amine such as triethylamine orN,N-diisopropylethylamine.

As an alternative, a reactive derivative of the carboxylic acid, e.g. ananhydride or acid chloride, may be used. Reaction with a reactivederivative such an anhydride is typically accomplished by stirring theamine and anhydride at room temperature in the presence of a base suchas pyridine.

Amines of the formula H₂N—Y—R³ can be obtained from commercial sourcesor can be prepared by any of a large number of standard syntheticmethods well known those skilled in the art, see for example seeAdvanced Organic Chemistry by Jerry March, 4^(th) Edition, John Wiley &Sons, 1992, and Organic Syntheses, Volumes 1-8, John Wiley, edited byJeremiah P. Freeman (ISBN: 0-471-31192-8), 1995, and see also themethods described in the experimental section below.

The nitro-pyrazole amide (XI) is reduced to give the corresponding4-amino-compound of the formula (XII). The reduction may be carried outby standard methods such as catalytic hydrogenation, for example in thepresence of palladium on carbon in a polar solvent such as ethanol ordimethylformamide at room temperature. As an alternative, reduction maybe effected using a reducing agent such as tin (II) chloride in ethanol,typically with heating, for example to the reflux temperature of thesolvent.

The 4-amino-pyrazole compound (XII) is then reacted with a carboxylicacid of the formula R¹—CO₂H, or a reactive derivative thereof, using themethods and conditions described above for the formation of the amide(XI), to give a compound of the formula (I).

Carboxylic acids of the formula R¹—CO₂H can be obtained commercially orcan be synthesised according to methods well known to the skilledperson, see for example Advanced Organic Chemistry and OrganicSyntheses, the details for which are given above.

Compounds of the formula (I) in which X is a group R¹-A-NR⁴, where A isa bond, can be prepared from the 4-amino compounds of the formula (XII)by a number of methods. Reductive amination with an appropriatelysubstituted aldehyde or ketone can be carried out in the presence ofvariety of reducing agents (see Advanced Organic Chemistry by JerryMarch, 4^(th) Edition, John Wiley & Sons, 1992, pp 898-900. For example,reductive amination can be carried out in the presence of sodiumtriacetoxyborohydride in the presence of an aprotic solvent such asdichloromethane at or near ambient temperatures.

Compounds in which X is a group R¹-A-NR⁴ where A is a bond can also beprepared by the reaction of the 4-amino pyrazole compound (XII) with acompound of the formula R¹—L in a nucleophilic displacement reactionwhere L is a leaving group such as a halogen.

In an alternative synthetic route, compounds of the formula (I) can beprepared by reaction of a compound of the formula (XIII) with a compoundof the formula R³—Y—NH₂. The reaction can be carried out using the amidecoupling conditions described above.

Compounds of the formula (I) where A is NH(C═O) can be prepared usingstandard methods for the synthesis of ureas. For example, such compoundscan be prepared by reacting an aminopyrazole compound of the formula(XII) with a suitably substituted phenylisocyanate in a polar solventsuch as DMF. The reaction is conveniently carried out at roomtemperature.

Compounds of the formula (I) where A is O(C═O) can be made usingstandard methods for the synthesis of carbamates, for example byreaction of an amino pyrazole compound of the formula (XII) with achloroformate derivative of the formula R¹—O—C(O)—Cl under conditionswell known to the skilled person.

Compounds of the formula (I), wherein A is SO₂, can be prepared fromamino-compounds of the formula (XII) by standard methods for theformation of sulphonamides. For example, compounds of the formula XII)can be reacted with sulphonyl chlorides of the formula R¹SO₂Cl oranhydrides of the formula (R¹SO₂)₂O. The reaction is typically carriedout in an aprotic solvent such as acetonitrile or a chlorinatedhydrocarbon (for example dichloromethane) in the presence of anon-interfering base such as a tertiary amine (e.g. triethylamine) orpyridine, or diisopropylethyl amine (Hunigs base). Alternatively, wherethe base is a liquid, as is the case with pyridine, the base itself maybe used as the solvent for the reaction.

Compounds wherein X is a 5- or 6-membered ring containing a carbon atomring member linked to the pyrazole group can be prepared by the sequenceof reactions set out in Scheme 2.

As shown in Scheme 2, an aldehyde (XIV) (in which X is a C-linked arylor heteroaryl group such as phenyl) is condensed with malononitrile togive the alkyne (XVI). The reaction is typically carried out in a polarsolvent such as ethanol in the presence of a base such as piperidine,usually with heating. The alkyne (XVI) is then reacted withtrimethylsilyldiazomethane in the presence an alkyl lithium such asbutyl lithium to give the 5-trimethylsilyl pyrazole-3-nitrile (XVII).The reaction is carried out in a dry aprotic solvent such as THF under aprotective atmosphere (e.g. nitrogen) at a reduced temperature (e.g.−78° C.).

The nitrile (XVII) is hydrolysed with an alkali metal hydroxide such aspotassium hydroxide to give the acid (XIX) and/or the amide (XVII).Where a mixture of acid and amide are formed, they may be separatedaccording to standard methods such as chromatography. The acid (XIX) canthen be coupled with an amine of the formula R³—Y—NH₂ under typicalamide coupling conditions of the type described above to give thecompound of the formula (I).

Alternatively, compounds of the formula (I) in which X is a C-linkedaryl or heteroaryl group such as phenyl can be prepared from compoundsof the formula (XX):

where “Hal” is a halogen such as chlorine, bromine or iodine, by meansof a Suzuki coupling reaction with the appropriate aryl or heteroarylboronate. The reaction can be carried out under typical Suzuki Couplingconditions in the presence of a palladium catalyst such asbis(tri-t-butylphosphine)palladium and a base (e.g. a carbonate such aspotassium carbonate). The reaction may be carried out in an aqueoussolvent system, for example aqueous ethanol, and the reaction mixture istypically subjected to heating, for example to a temperature in excessof 100° C.

Compounds of the formula (XX) can be prepared from amino-pyrazolecompounds of the formula (XII) by means of the Sandmeyer reaction (seeAdvanced Organic Chemistry, 4^(th) edition, by Jerry March, John Wiley &Sons, 1992, page 723) in which the amino group is converted to adiazonium group by reaction with nitrous acid, and the diazoniumcompound is then reacted with a copper (I) halide such as Cu(I)Cl orCu(I)I.

Once formed, one compound of the formula (I) may be transformed intoanother compound of the formula (I) using standard chemistry procedureswell known in the art. For examples of functional groupinterconversions, see for example, Fiesers' Reagents for OrganicSynthesis, Volumes 1-17, John Wiley, edited by Mary Fieser (ISBN:0-471-58283-2), and Organic Syntheses, Volumes 1-8, John Wiley, editedby Jeremiah P. Freeman (ISBN: 0-471-31192-8), 1995.

The starting materials for the synthetic routes shown in the Schemesabove, e.g. the pyrazoles of formula (X), can either be obtainedcommercially or can be prepared by methods known to those skilled in theart. They can be obtained using known methods e.g. from ketones, such asin a process described in EP308020 (Merck), or the methods discussed bySchmidt in Helv. Chim. Acta., 1956, 39, 986-991 and Helv. Chim. Acta.,1958, 41, 306-309. Alternatively they can be obtained by conversion of acommercially available pyrazole, for example those containing halogen,nitro, ester, or amide functionalities, to pyrazoles containing thedesired functionality by standard methods known to a person skilled inthe art. For example, in 3-carboxy-4-nitropyrazole, the nitro group canbe reduced to an amine by standard methods.4-Nitro-pyrazole-3-carboxylic acid (XII) can either be obtainedcommercially or can be prepared by nitration of the corresponding4-unsubstituted pyrazole carboxy compound, and pyrazoles containing ahalogen, may be utilized in coupling reactions with tin or palladiumchemistry.

Protecting Groups

In many of the reactions described above, it may be necessary to protectone or more groups to prevent reaction from taking place at anundesirable location on the molecule. Examples of protecting groups, andmethods of protecting and deprotecting functional groups, can be foundin Protective Groups in Organic Synthesis (T. Green and P. Wuts; 3rdEdition; John Wiley and Sons, 1999).

A hydroxy group may be protected, for example, as an ether (—OR) or anester (—OC(═O)R), for example, as: a t-butyl ether; a tetrahydropyranyl(THP) ether; a benzyl, benzhydryl (diphenylmethyl), or trityl(triphenylmethyl)ether; a trimethylsilyl or t-butyldimethylsilyl ether;or an acetyl ester (—OC(═O)CH₃, —OAc).

An aldehyde or ketone group may be protected, for example, as an acetal(R—CH(OR)₂) or ketal (R₂C(OR)₂), respectively, in which the carbonylgroup (>C═O) is converted to a diether (>C(OR)₂), by reaction with, forexample, a primary alcohol. The aldehyde or ketone group is readilyregenerated by hydrolysis using a large excess of water in the presenceof acid.

An amine group may be protected, for example, as an amide (—NRCO—R) or aurethane (—NRCO—OR), for example, as: a methyl amide (—NHCO—CH₃); abenzyloxy amide (—NHCO—OCH₂C₆H₅, —NH-Cbz or NH—Z); as a t-butoxy amide(—NHCO—OC(CH₃)₃, —NH-Boc); a 2-biphenyl-2-propoxy amide(—NHCO—OC(CH₃)₂C₆H₄C₆H₅, —NH-Bpoc), as a 9-fluorenylmethoxy amide(—NH-Fmoc), as a 6-nitroveratryloxy amide (—NH-Nvoc), as a2-trimethylsilylethyloxy amide (—NH-Teoc), as a 2,2,2-trichloroethyloxyamide (—NH-Troc), as an allyloxy amide (—NH-Alloc), or as a2(-phenylsulphonyl)ethyloxy amide (—NH-Psec).

For example, in Scheme 1 above, when the moiety R³ in the amine H₂N—Y—R³contains a second amino group, such as a cyclic amino group (e.g. apiperidine or pyrrolidine group), the second amino group can beprotected by means of a protecting group as hereinbefore defined, onepreferred group being the tert-butyloxycarbonyl (Boc) group. Where nosubsequent modification of the second amino group is required, theprotecting group can be carried through the reaction sequence to give anN-protected form of a compound of the formula (I) which can then bede-protected by standard methods (e.g. treatment with acid in the caseof the Boc group) to give the compound of formula (I).

Other protecting groups for amines, such as cyclic amines andheterocyclic N—H groups, include toluenesulphonyl (tosyl) andmethanesulphonyl (mesyl) groups, benzyl groups such as apara-methoxybenzyl (PMB) group and tetrahydropyranyl (THP) groups.

A carboxylic acid group may be protected as an ester for example, as: anC₁₋₇ alkyl ester (e.g., a methyl ester; a t-butyl ester); aC₁₋₇haloalkyl ester (e.g., a C₁₋₇ trihaloalkyl ester); a triC₁₋₇alkylsilyl-C₁₋₇alkyl ester; or a C₅₋₂₀ aryl-C₁₋₇ alkyl ester (e.g., abenzyl ester; a nitrobenzyl ester); or as an amide, for example, as amethyl amide. A thiol group may be protected, for example, as athioether (—SR), for example, as: a benzyl thioether; an acetamidomethylether (—S—CH₂NHC(═O)CH₃).

Isolation and Purification of the Compounds of the Invention

The compounds of the invention can be isolated and purified according tostandard techniques well known to the person skilled in the art. Onetechnique of particular usefulness in purifying the compounds ispreparative liquid chromatography using mass spectrometry as a means ofdetecting the purified compounds emerging from the chromatographycolumn.

Preparative LC-MS is a standard and effective method used for thepurification of small organic molecules such as the compounds describedherein. The methods for the liquid chromatography (LC) and massspectrometry (MS) can be varied to provide better separation of thecrude materials and improved detection of the samples by MS.Optimisation of the preparative gradient LC method will involve varyingcolumns, volatile eluents and modifiers, and gradients. Methods are wellknown in the art for optimising preparative LC-MS methods and then usingthem to purify compounds. Such methods are described in Rosentreter U,Huber U.; Optimal fraction collecting in preparative LC/MS; J CombChem.; 2004; 6(2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z,Lindsley C., Development of a custom high-throughput preparative liquidchromatography/mass spectrometer platform for the preparativepurification and analytical analysis of compound libraries; J CombChem.; 2003; 5(3); 322-9.

An example of such a system for purifying compounds via preparativeLC-MS is described below in the Examples section of this application(under the heading “Mass Directed Purification LC-MS System”). However,it will be appreciated that alternative systems and methods to thosedescribed could be used. In particular, normal phase preparative LCbased methods might be used in place of the reverse phase methodsdescribed here. Most preparative LC-MS systems utilise reverse phase LCand volatile acidic modifiers, since the approach is very effective forthe purification of small molecules and because the eluents arecompatible with positive ion electrospray mass spectrometry. Employingother chromatographic solutions e.g. normal phase LC, alternativelybuffered mobile phase, basic modifiers etc as outlined in the analyticalmethods described below could alternatively be used to purify thecompounds.

Pharmaceutical Formulations

While it is possible for the active compound to be administered alone,it is preferable to present it as a pharmaceutical composition (e.g.formulation) comprising at least one active compound of the inventiontogether with one or more pharmaceutically acceptable carriers,adjuvants, excipients, diluents, fillers, buffers, stabilisers,preservatives, lubricants, or other materials well known to thoseskilled in the art and optionally other therapeutic or prophylacticagents.

Thus, the present invention further provides pharmaceuticalcompositions, as defined above, and methods of making a pharmaceuticalcomposition comprising admixing at least one active compound, as definedabove, together with one or more pharmaceutically acceptable carriers,excipients, buffers, adjuvants, stabilizers, or other materials, asdescribed herein.

The term “pharmaceutically acceptable” as used herein pertains tocompounds, materials, compositions, and/or dosage forms which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of a subject (e.g. human) without excessive toxicity,irritation, allergic response, or other problem or complication,commensurate with a reasonable benefit/risk ratio. Each carrier,excipient, etc. must also be “acceptable” in the sense of beingcompatible with the other ingredients of the formulation.

Accordingly, in a further aspect, the invention provides compounds ofthe formula (O) and sub-groups thereof such as formulae (I⁰), (I), (Ia),(Ib), (II), (III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or(VIII) and sub-groups thereof as defined herein in the form ofpharmaceutical compositions.

The pharmaceutical compositions can be in any form suitable for oral,parenteral, topical, intranasal, ophthalmic, otic, rectal,intra-vaginal, or transdermal administration. Where the compositions areintended for parenteral administration, they can be formulated forintravenous, intramuscular, intraperitoneal, subcutaneous administrationor for direct delivery into a target organ or tissue by injection,infusion or other means of delivery.

In one preferred embodiment of the invention, the pharmaceuticalcomposition is in a form suitable for i.v. administration, for exampleby injection or infusion.

In another preferred embodiment, the pharmaceutical composition is in aform suitable for sub-cutaneous (s.c.) administration.

Pharmaceutical dosage forms suitable for oral administration includetablets, capsules, caplets, pills, lozenges, syrups, solutions, powders,granules, elixirs and suspensions, sublingual tablets, wafers or patchesand buccal patches.

Pharmaceutical compositions containing compounds of the formula (I) canbe formulated in accordance with known techniques, see for example,Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton,Pa., USA.

Thus, tablet compositions can contain a unit dosage of active compoundtogether with an inert diluent or carrier such as a sugar or sugaralcohol, eg; lactose, sucrose, sorbitol or mannitol; and/or a non-sugarderived diluent such as sodium carbonate, calcium phosphate, calciumcarbonate, or a cellulose or derivative thereof such as methylcellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and starchessuch as corn starch. Tablets may also contain such standard ingredientsas binding and granulating agents such as polyvinylpyrrolidone,disintegrants (e.g. swellable crosslinked polymers such as crosslinkedcarboxymethylcellulose), lubricating agents (e.g. stearates),preservatives (e.g. parabens), antioxidants (e.g. BHT), buffering agents(for example phosphate or citrate buffers), and effervescent agents suchas citrate/bicarbonate mixtures. Such excipients are well known and donot need to be discussed in detail here.

Capsule formulations may be of the hard gelatin or soft gelatin varietyand can contain the active component in solid, semi-solid, or liquidform. Gelatin capsules can be formed from animal gelatin or synthetic orplant derived equivalents thereof.

The solid dosage forms (eg; tablets, capsules etc.) can be coated orun-coated, but typically have a coating, for example a protective filmcoating (e.g. a wax or varnish) or a release controlling coating. Thecoating (e.g. a Eudragit™ type polymer) can be designed to release theactive component at a desired location within the gastro-intestinaltract. Thus, the coating can be selected so as to degrade under certainpH conditions within the gastrointestinal tract, thereby selectivelyrelease the compound in the stomach or in the ileum or duodenum.

Instead of, or in addition to, a coating, the drug can be presented in asolid matrix comprising a release controlling agent, for example arelease delaying agent which may be adapted to selectively release thecompound under conditions of varying acidity or alkalinity in thegastrointestinal tract. Alternatively, the matrix material or releaseretarding coating can take the form of an erodible polymer (e.g. amaleic anhydride polymer) which is substantially continuously eroded asthe dosage form passes through the gastrointestinal tract. As a furtheralternative, the active compound can be formulated in a delivery systemthat provides osmotic control of the release of the compound. Osmoticrelease and other delayed release or sustained release formulations maybe prepared in accordance with methods well known to those skilled inthe art.

Compositions for topical use include ointments, creams, sprays, patches,gels, liquid drops and inserts (for example intraocular inserts). Suchcompositions can be formulated in accordance with known methods.

Compositions for parenteral administration are typically presented assterile aqueous or oily solutions or fine suspensions, or may beprovided in finely divided sterile powder form for making upextemporaneously with sterile water for injection.

Examples of formulations for rectal or intra-vaginal administrationinclude pessaries and suppositories which may be, for example, formedfrom a shaped moldable or waxy material containing the active compound.

Compositions for administration by inhalation may take the form ofinhalable powder compositions or liquid or powder sprays, and can beadministrated in standard form using powder inhaler devices or aerosoldispensing devices. Such devices are well known. For administration byinhalation, the powdered formulations typically comprise the activecompound together with an inert solid powdered diluent such as lactose.

The compounds of the inventions will generally be presented in unitdosage form and, as such, will typically contain sufficient compound toprovide a desired level of biological activity. For example, aformulation intended for oral administration may contain from 0.1milligrams to 2 grams of active ingredient, more usually from 10milligrams to 1 gram, for example, 50 milligrams to 500 milligrams.

The active compound will be administered to a patient in need thereof(for example a human or animal patient) in an amount sufficient toachieve the desired therapeutic effect.

Methods of Treatment

It is envisaged that the compounds of the formula (0) and sub-groupsthereof such as formulae (I⁰), (I), (Ia), (Ib), (II), (III), (IV),(IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereofas defined herein will be useful in the prophylaxis or treatment of arange of disease states or conditions mediated by cyclin dependentkinases. Examples of such disease states and conditions are set outabove.

The compounds are generally administered to a subject in need of suchadministration, for example a human or animal patient, preferably ahuman.

The compounds will typically be administered in amounts that aretherapeutically or prophylactically useful and which generally arenon-toxic. However, in certain situations (for example in the case oflife threatening diseases), the benefits of administering a compound ofthe formula (I) may outweigh the disadvantages of any toxic effects orside effects, in which case it may be considered desirable to administercompounds in amounts that are associated with a degree of toxicity.

The compounds may be administered over a prolonged term to maintainbeneficial therapeutic effects or may be administered for a short periodonly. Alternatively they may be administered in a pulsatile orcontinuous manner.

A typical daily dose of the compound can be in the range from 100picograms to 100 milligrams per kilogram of body weight, more typically5 nanograms to 25 milligrams per kilogram of bodyweight, and moreusually 10 nanograms to 15 milligrams per kilogram (e.g. 10 nanograms to10 milligrams) per kilogram of bodyweight although higher or lower dosesmay be administered where required. Ultimately, the quantity of compoundadministered and the type of composition used will be commensurate withthe nature of the disease or physiological condition being treated andwill be at the discretion of the physician.

The compounds of the formula (I) can be administered as the soletherapeutic agent or they can be administered in combination therapywith one of more other compounds for treatment of a particular diseasestate, for example a neoplastic disease such as a cancer as hereinbeforedefined. Examples of other therapeutic agents that may be administeredtogether (whether concurrently or at different time intervals) with thecompounds of the formula (I) include but are not limited totopoisomerase inhibitors, alkylating agents, antimetabolites, DNAbinders and microtubule inhibitors (tubulin targeting agents), such ascisplatin, cyclophosphamide, doxorubicin, irinotecan, fludarabine, 5FU,taxanes, mitomycin C, or radiotherapy. Alternatively, the compounds ofthe formula (I) can be administered in a combination therapy withmonoclonal antibodies or signal transduction inhibitors. For the case ofCDK inhibitors combined with other therapies, the two or more treatmentsmay be given in individually varying dose schedules and via differentroutes.

Where the compound of the formula (I) is administered in combinationtherapy with one, two, three, four or more other therapeutic agents(preferably one or two, more preferably one), the compounds can beadministered simultaneously or sequentially. When administeredsequentially, they can be administered at closely spaced intervals (forexample over a period of 5-10 minutes) or at longer intervals (forexample 1, 2, 3, 4 or more hours apart, or even longer periods apartwhere required), the precise dosage regimen being commensurate with theproperties of the therapeutic agent(s).

The compounds of the invention may also be administered in conjunctionwith non-chemotherapeutic treatments such as radiotherapy, photodynamictherapy, gene therapy; surgery and controlled diets.

For use in combination therapy with another chemotherapeutic agent, thecompound of the formula (I) and one, two, three, four or more othertherapeutic agents can be, for example, formulated together in a dosageform containing two, three, four or more therapeutic agents. In analternative, the individual therapeutic agents may be formulatedseparately and presented together in the form of a kit, optionally withinstructions for their use.

A person skilled in the art would know through their common generalknowledge the dosing regimes and combination therapies to use.

Methods of Diagnosis

Prior to administration of a compound of the formula (I), a patient maybe screened to determine whether a disease or condition from which thepatient is or may be suffering is one which would be susceptible totreatment with a compound having activity against cyclin dependentkinases.

For example, a biological sample taken from a patient may be analysed todetermine whether a condition or disease, such as cancer, that thepatient is or may be suffering from is one which is characterised by agenetic abnormality or abnormal protein expression which leads toover-activation of CDKs or to sensitisation of a pathway to normal CDKactivity. Examples of such abnormalities that result in activation orsensitisation of the CDK2 signal include up-regulation of cyclin E,(Harwell R M, Mull B B, Porter D C, Keyomarsi K.; J Biol. Chem. 2004Mar. 26; 279(13):12695-705) or loss of p21 or p27, or presence of CDC4variants (Rajagopalan H, Jallepalli P V, Rago C, Velculescu V E, KinzlerK W, Vogelstein B, Lengauer C.; Nature. 2004 Mar. 4; 428(6978):77-81).The term up-regulation includes elevated expression or over-expression,including gene amplification (i.e. multiple gene copies) and increasedexpression by a transcriptional effect, and hyperactivity andactivation, including activation by mutations. Thus, the patient may besubjected to a diagnostic test to detect a marker characteristic ofup-regulation of cyclin E, or loss of p21 or p27, or presence of CDC4variants. The term diagnosis includes screening. By marker we includegenetic markers including, for example, the measurement of DNAcomposition to identify mutations of CDC4. The term marker also includesmarkers which are characteristic of up regulation of cyclin E, includingenzyme activity, enzyme levels, enzyme state (e.g. phosphorylated ornot) and mRNA levels of the aforementioned proteins.

Tumours with upregulation of cyclin E, or loss of p21 or p27 may beparticularly sensitive to CDK inhibitors. Tumours may preferentially bescreened for upregulation of cyclin E, or loss of p21 or p27 prior totreatment. Thus, the patient may be subjected to a diagnostic test todetect a marker characteristic of up-regulation of cyclin E, or loss ofp21 or p27. The diagnostic tests are typically conducted on a biologicalsample selected from tumour biopsy samples, blood samples (isolation andenrichment of shed tumour cells), stool biopsies, sputum, chromosomeanalysis, pleural fluid, peritoneal fluid, or urine.

It has been found, Rajagopalan et al (Nature. 2004 Mar. 4;428(6978):77-81), that there were mutations present in CDC4 (also knownas Fbw7 or Archipelago) in human colorectal cancers and endometrialcancers (Spruck et al, Cancer Res. 2002 Aug. 15; 62(16):4535-9).Identification of individual carrying a mutation in CDC4 may mean thatthe patient would be particularly suitable for treatment with a CDKinhibitor. Tumours may preferentially be screened for presence of a CDC4variant prior to treatment. The screening process will typically involvedirect sequencing, oligonucleotide microarray analysis, or a mutantspecific antibody.

Methods of identification and analysis of mutations and up-regulation ofproteins are known to a person skilled in the art. Screening methodscould include, but are not limited to, standard methods such asreverse-transcriptase polymerase chain reaction (RT-PCR) or in-situhybridisation.

In screening by RT-PCR, the level of mRNA in the tumour is assessed bycreating a cDNA copy of the mRNA followed by amplification of the cDNAby PCR. Methods of PCR amplification, the selection of primers, andconditions for amplification, are known to a person skilled in the art.Nucleic acid manipulations and PCR are carried out by standard methods,as described for example in Ausubel, F. M. et al., eds. CurrentProtocols in Molecular Biology, 2004, John Wiley & Sons Inc., or Innis,M. A. et-al., eds. PCR Protocols: a guide to methods and applications,1990, Academic Press, San Diego. Reactions and manipulations involvingnucleic acid techniques are also described in Sambrook et al., 2001,3^(rd) Ed, Molecular Cloning: A Laboratory Manual, Cold Spring HarborLaboratory Press. Alternatively a commercially available kit for RT-PCR(for example Roche Molecular Biochemicals) may be used, or methodologyas set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531;5,192,659, 5,272,057, 5,882,864, and 6,218,529 and incorporated hereinby reference.

An example of an in-situ hybridisation technique for assessing mRNAexpression would be fluorescence in-situ hybridisation (FISH) (seeAngerer, 1987 Meth. Enzymol., 152: 649).

Generally, in situ hybridization comprises the following major steps:(1) fixation of tissue to be analyzed; (2) prehybridization treatment ofthe sample to increase accessibility of target nucleic acid, and toreduce nonspecific binding; (3) hybridization of the mixture of nucleicacids to the nucleic acid in the biological structure or tissue; (4)post-hybridization washes to remove nucleic acid fragments not bound inthe hybridization, and (5) detection of the hybridized nucleic acidfragments. The probes used in such applications are typically labeled,for example, with radioisotopes or fluorescent reporters. Preferredprobes are sufficiently long, for example, from about 50, 100, or 200nucleotides to about 1000 or more nucleotides, to enable specifichybridization with the target nucleic acid(s) under stringentconditions. Standard methods for carrying out FISH are described inAusubel, F. M. et al., eds. Current Protocols in Molecular Biology,2004, John Wiley & Sons Inc and Fluorescence In Situ Hybridization:Technical Overview by John M. S. Bartlett in Molecular Diagnosis ofCancer, Methods and Protocols, 2nd ed.; ISBN: 1-59259-760-2; March 2004,pps. 077-088; Series: Methods in Molecular Medicine.

Alternatively, the protein products expressed from the mRNAs may beassayed by immunohistochemistry of tumour samples, solid phaseimmunoassay with microtiter plates, Western blotting, 2-dimensionalSDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry and othermethods known in the art for detection of specific proteins. Detectionmethods would include the use of site specific antibodies. The skilledperson will recognize that all such well-known techniques for detectionof upregulation of cyclin E, or loss of p21 or p2′7, or detection ofCDC4 variants could be applicable in the present case.

Therefore all of these techniques could also be used to identify tumoursparticularly suitable for treatment with CDK inhibitors. Patients withmantle cell lymphoma (MCL) could be selected for treatment with a CDKinhibitor using diagnostic tests outlined herein. MCL is a distinctclinicopathologic entity of non-Hodgkin's lymphoma, characterized byproliferation of small to medium-sized lymphocytes with co-expression ofCD5 and CD20, an aggressive and incurable clinical course, and frequentt(11;14)(q13;q32) translocation. Over-expression of cyclin D1 mRNA,found in mantle cell lymphoma (MCL), is a critical diagnostic marker.Yatabe et al (Blood. 2000 Apr. 1; 95(7):2253-61) proposed that cyclinD1-positivity should be included as one of the standard criteria forMCL, and that innovative therapies for this incurable disease should beexplored on the basis of the new criteria. Jones et al (J Mol Diagn.2004 May; 6(2):84-9) developed a real-time, quantitative, reversetranscription PCR assay for cyclin D1 (CCND1) expression to aid in thediagnosis of mantle cell lymphoma (MCL). Howe et al (Clin Chem. 2004January; 50(1):80-7) used real-time quantitative RT-PCR to evaluatecyclin D1 mRNA expression and found that quantitative RT-PCR for cyclinD1 mRNA normalized to CD19 mRNA can be used in the diagnosis of MCL inblood, marrow, and tissue. Alternatively, patients with breast cancercould be selected for treatment with a CDK inhibitor using diagnostictests outline above. Tumour cells commonly overexpress cyclin E and ithas been shown that cyclin E is over-expressed in breast cancer (Harwellet al, Cancer Res, 2000, 60, 481-489). Therefore breast cancer may inparticular be treated with a CDK inhibitor.

Antifungal Use

In a further aspect, the invention provides the use of the compounds ofthe formulae (0), (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va),(Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereof as definedherein as antifungal agents.

The compounds may be used in animal medicine (for example in thetreatment of mammals such as humans), or in the treatment of plants(e.g. in agriculture and horticulture), or as general antifungal agents,for example as preservatives and disinfectants.

In one embodiment, the invention provides a compound of the formula (0)and sub-groups thereof such as formulae (I⁰), (I), (Ia), (Ib), (II),(III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) andsub-groups thereof as defined herein for use in the prophylaxis ortreatment of a fungal infection in a mammal such as a human.

Also provided is the use of a compound of the formula (0) and sub-groupsthereof such as formulae (I⁰), (I), (Ia), (Ib), (II), (III), (IV),(IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) and sub-groups thereofas defined herein for the manufacture of a medicament for use in theprophylaxis or treatment of a fungal infection in a mammal such as ahuman.

For example, compounds of the invention may be administered to humanpatients suffering from, or at risk of infection by, topical fungalinfections caused by among other organisms, species of Candida,Trichophyton, Microsporum or Epidermophyton, or in mucosal infectionscaused by Candida albicans (e.g. thrush and vaginal candidiasis). Thecompounds of the invention can also be administered for the treatment orprophylaxis of systemic fungal infections caused by, for example,Candida albicans, Cryptococcus neoformans, Aspergillus flavus,Aspergillus fumigatus, Coccidiodies, Paracoccidioides, Histoplasma orBlastomyces.

In another aspect, the invention provides an antifungal composition foragricultural (including horticultural) use, comprising a compound of theformula (I⁰) and sub-groups thereof such as formulae (I), (Ia), (Ib),(II), (III), (IV), (V), (VI) and (VII) as hereinbefore defined togetherwith an agriculturally acceptable diluent or carrier.

The invention further provides a method of treating an animal (includinga mammal such as a human), plant or seed having a fungal infection,which comprises treating said animal, plant or seed, or the locus ofsaid plant or seed, with an effective amount of a compound of theformula (I⁰) and sub-groups thereof such as formulae (I), (Ia), (Ib),(II), (III), (IV), (V), (VI) and (VII) as hereinbefore defined.

The invention also provides a method of treating a fungal infection in aplant or seed which comprises treating the plant or seed with anantifungally effective amount of a fungicidal composition ashereinbefore defined.

Differential screening assays may be used to select for those compoundsof the present invention with specificity for non-human CDK enzymes.Compounds which act specifically on the CDK enzymes of eukaryoticpathogens can be used as anti-fungal or anti-parasitic agents.Inhibitors of the Candida CDK kinase, CKSI, can be used in the treatmentof candidiasis. Antifungal agents can be used against infections of thetype hereinbefore defined, or opportunistic infections that commonlyoccur in debilitated and immunosuppressed patients such as patients withleukemias and lymphomas, people who are receiving immunosuppressivetherapy, and patients with predisposing conditions such as diabetesmellitus or AIDS, as well as for non-immunosuppressed patients.

Assays described in the art can be used to screen for agents which maybe useful for inhibiting at least one fungus implicated in mycosis suchas candidiasis, aspergillosis, mucormycosis, blastomycosis,geotrichosis, cryptococcosis, chromoblastomycosis, coccidiodomycosis,conidiosporosis, histoplasmosis, maduromycosis, rhinosporidosis,nocaidiosis, para-actinomycosis, penicilliosis, monoliasis, orsporotrichosis. The differential screening assays can be used toidentify anti-fungal agents which may have therapeutic value in thetreatment of aspergillosis by making use of the CDK genes cloned fromyeast such as Aspergillus fumigatus, Aspergillus flavus, Aspergillusniger, Aspergillus nidulans, or Aspergillus terreus, or where themycotic infection is mucon-nycosis, the CDK assay can be derived fromyeast such as Rhizopus arrhizus, Rhizopus oryzae, Absidia corymbifera,Absidia ramosa, or Mucor pusillus. Sources of other CDK enzymes includethe pathogen Pneumocystis carinii.

By way of example, in vitro evaluation of the antifungal activity of thecompounds can be performed by determining the minimum inhibitoryconcentration (M.I.C.) which is the concentration of the test compounds,in a suitable medium, at which growth of the particular microorganismfails to occur. In practice, a series of agar plates, each having thetest compound incorporated at a particular concentration is inoculatedwith a standard culture of, for example, Candida albicans and each plateis then incubated for an appropriate period at 37° C. The plates arethen examined for the presence or absence of growth of the fungus andthe appropriate M.I.C. value is noted

The in vivo evaluation of the compounds can be carried out at a seriesof dose levels by intraperitoneal or intravenous injection or by oraladministration, to mice that have been inoculated with a fungus, e.g., astrain of Candida albicans or Aspergillus flavus. The activity of thecompounds can be assessed on the basis of the survival of a treatedgroup of mice after the death of an untreated group of mice. Theactivity may be measured in terms of the dose level at which thecompound provides 50% protection against the lethal effect of theinfection (PD₅₀).

For human antifungal use, the compounds can be administered alone or inadmixture with a pharmaceutical carrier selected in accordance with theintended route of administration and standard pharmaceutical practice.Thus, for example, they may be administered orally, parenterally,intravenously, intramuscularly or subcutaneously by means of theformulations described above in the section headed “PharmaceuticalFormulations”.

For oral and parenteral administration to human patients, the dailydosage level of the antifungal compounds of the invention can be from0.01 to 10 mg/kg (in divided doses), depending on inter alia the potencyof the compounds when administered by either the oral or parenteralroute. Tablets or capsules of the compounds may contain, for example,from 5 mg. to 0.5 g of active compound for administration singly or twoor more at a time as appropriate. The physician in any event willdetermine the actual dosage (effective amount) which will be mostsuitable for an individual patient and it will vary with the age, weightand response of the particular patient.

Alternatively, the antifungal compounds can be administered in the formof a suppository or pessary, or they may be applied topically in theform of a lotion, solution, cream, ointment or dusting powder. Forexample, they can be incorporated into a cream consisting of an aqueousemulsion of polyethylene glycols or liquid paraffin; or they can beincorporated, at a concentration between 1 and 10%, into an ointmentconsisting of a white wax or white soft paraffin base together with suchstabilizers and preservatives as may be required.

In addition to the therapeutic uses described above, anti-fungal agentsdeveloped with such differential screening assays can be used, forexample, as preservatives in foodstuff, feed supplement for promotingweight gain in livestock, or in disinfectant formulations for treatmentof non-living matter, e.g., for decontaminating hospital equipment androoms. In similar fashion, side by side comparison of inhibition of amammalian CDK and an insect CDK, such as the Drosophilia CDK5 gene(Hellmich et al. (1994) FEBS Lett 356:317-21), will permit selectionamongst the compounds herein of inhibitors which discriminate betweenthe human/mammalian and insect enzymes. Accordingly, the presentinvention expressly contemplates the use and formulations of thecompounds of the invention in insecticides, such as for use inmanagement of insects like the fruit fly.

In yet another embodiment, certain of the subject CDK inhibitors can beselected on the basis of inhibitory specificity for plant CDK's relativeto the mammalian enzyme. For example, a plant CDK can be disposed in adifferential screen with one or more of the human enzymes to selectthose compounds of greatest selectivity for inhibiting the plant enzyme.Thus, the present invention specifically contemplates formulations ofthe subject CDK inhibitors for agricultural applications, such as in theform of a defoliant or the like.

For agricultural and horticultural purposes the compounds of theinvention may be used in the form of a composition formulated asappropriate to the particular use and intended purpose. Thus thecompounds may be applied in the form of dusting powders, or granules,seed dressings, aqueous solutions, dispersions or emulsions, dips,sprays, aerosols or smokes. Compositions may also be supplied in theform of dispersible powders, granules or grains, or concentrates fordilution prior to use. Such compositions may contain such conventionalcarriers, diluents or adjuvants as are known and acceptable inagriculture and horticulture and they are manufactured in accordancewith conventional procedures. The compositions may also incorporateother active ingredients, for example, compounds having herbicidal orinsecticidal activity or a further fungicide. The compounds andcompositions can be applied in a number of ways, for example they can beapplied directly to the plant foliage, stems, branches, seeds or rootsor to the soil or other growing medium, and they may be used not only toeradicate disease, but also prophylactically to protect the plants orseeds from attack. By way of example, the compositions may contain from0.01 to 1 wt. % of the active ingredient. For field use, likelyapplication rates of the active ingredient may be from 50 to 5000g/hectare.

The invention also contemplates the use of the compounds of the formula(0) and sub-groups thereof such as formulae (I⁰), (I), (Ia), (Ib), (II),(III), (IV), (IVa), (Va), (Vb), (VIa), (VIb), (VII) or (VIII) andsub-groups thereof as defined herein in the control of wood decayingfungi and in the treatment of soil where plants grow, paddy fields forseedlings, or water for perfusion. Also contemplated by the invention isthe use of the compounds of the formula (0) and sub-groups thereof suchas formulae (I⁰), (I), (Ia), (Ib), (II), (III), (IV), (IVa), (Va), (Vb),(VIa), (VIb), (VII) or (VIII) and sub-groups thereof as defined hereinto protect stored grain and other non-plant loci from fungalinfestation.

EXAMPLES

The invention will now be illustrated, but not limited, by reference tothe specific embodiments described in the following examples.

In the examples, the compounds prepared were characterised by liquidchromatography and mass spectroscopy (LC-MS) using the system andoperating conditions set out below. Where chlorine is present and asingle mass is quoted, the mass quoted for the compound is for ³⁵Cl. Thetwo systems were equipped with identical chromatography columns and wereset up to run under the same operating conditions. The operatingconditions used are also described below. In the examples, the retentiontimes are given in minutes.

Platform System System: Waters 2790/Platform LC Mass Spec Detector:Micromass Platform LC PDA Detector: Waters 996 PDA AnalyticalConditions: Eluent A: 5% CH3CN in 95% H₂O (0.1% Formic Acid) Eluent B:CH₃CN (0.1% Formic Acid)

Gradient: 10-95% eluent BFlow: 1.2 ml/min

Column: Synergi 4 μm Max-RP C₁₂, 80A, 50×4.6 mm (Phenomenex) MSConditions:

Capillary voltage: 3.5 kVCone voltage: 30 V

Source Temperature: 120° C. FractionLynx System

System: Waters FractionLynx (dual analytical/prep)

Mass Spec Detector: Waters-Micromass ZQ PDA Detector: Waters 2996 PDAAnalytical Conditions: Eluent A: H₂O (0.1% Formic Acid) Eluent B: CH₃CN(0.1% Formic Acid)

Gradient: 5-95% eluent BFlow: 1.5 ml/min

Column: Synergi 4 μm Max-RP C₁₂, 80A, 50×4.6 mm (Phenomenex) MSConditions:

Capillary voltage: 3.5 kVCone voltage: 30 V

Source Temperature: 120° C. Desolvation Temperature: 300° C. AnalyticalLC-MS System

Several systems were used, as described below, and these were equippedwith were set up to run under closely similar operating conditions. Theoperating conditions used are also described below.

HPLC System: Waters 2795 Mass Spec Detector: Micromass Platform LC PDADetector: Waters 2996 PDA Acidic Analytical Conditions: Eluent A: H₂O(0.1% Formic Acid) Eluent B: CH₃CN (0.1% Formic Acid)

Gradient: 5-95% eluent B over 3.5 minutesFlow: 0.8 ml/min

Column: Phenomenex Synergi 4μ MAX-RP 80A, 2.0×50 mm Basic AnalyticalConditions:

Eluent A: H₂O (10 mM NH₄HCO₃ buffer adjusted to pH=9.5 with NH₄OH)

Eluent B: CH₃CN

Gradient: 05-95% eluent B over 3.5 minutesFlow: 0.8 ml/min

Column: Thermo Hypersil-Keystone BetaBasic-18 5 μm 2.1×50 mm

or

Column: Phenomenex Luna C18(2) 5 μm 2.0×50 mm Polar AnalyticalConditions: Eluent A: H₂O (0.1% Formic Acid) Eluent B: CH₃CN (0.1%Formic Acid)

Gradient: 00-50% eluent B over 3 minutesFlow: 0.8 ml/min

Column: Thermo Hypersil-Keystone HyPurity Aquastar, 5μ, 2.1×50 mm

or

Column: Phenomenex Synergi 4μ MAX-RP 80A, 2.0×50 mm or Longer AnalyticalConditions: Eluent A: H₂O (0.1% Formic Acid) Eluent B: CH₃CN (0.1%Formic Acid)

Gradient: 05-95% eluent B over 15 minutesFlow: 0.4 ml/min

Column: Phenomenex Synergi 4μ MAX-RP 80A, 2.0×150 mm MS Conditions:

Capillary voltage: 3.6 kVCone voltage: 30 V

Source Temperature: 120° C. Scan Range: 165-700 amu Ionisation Mode:ElectroSpray Positive or

-   -   ElectroSpray Negative or    -   ElectroSpray Positive & Negative

Mass Directed Purification LC-MS System

The following preparative chromatography systems can be used to purifythe compounds of the invention.

Hardware:

Waters Fractionlynx system:

2767 Dual Autosampler/Fraction Collector

2525 preparative pumpCFO (column fluidic organiser) for column selectionRMA (Waters reagent manager) as make up pump

Waters ZQ Mass Spectrometer

Waters 2996 Photo Diode Array detector

Software: Masslynx 4.0

Columns:

1. Low pH chromatography: Phenomenex Synergy MAX-RP, 10μ, 150×15 mm(alternatively used same column type with 100×21.2 mm dimensions).2. High pH chromatography: Phenomenex Luna C18 (2), 10μ, 100×21.2 mm(alternatively used Thermo Hypersil Keystone BetaBasic C18, 5μ, 100×21.2mm)

Eluents:

1. Low pH chromatography:

Solvent A: H₂O+0.1% Formic Acid, pH 1.5 Solvent B: CH₃CN+0.1% FormicAcid

2. High pH chromatography:

Solvent A: H₂O+10 mM NH₄HCO₃+NH₄OH, pH 9.5 Solvent B: CH₃CN

3. Make up solvent: MeOH+0.1% formic acid (for both chromatography type)

Methods:

Prior to using preparative chromatography to isolate and purify theproduct compounds, analytical LC-MS (see above) can first be used todetermine the most appropriate conditions for preparativechromatography. A typical routine is to run an analytical LC-MS usingthe type of chromatography (low or high pH) most suited for compoundstructure. Once the analytical trace shows good chromatography, asuitable preparative method of the same type can be chosen. Typicalrunning condition for both low and high pH chromatography methods are:

Flow rate: 24 ml/minGradient: Generally all gradients have an initial 0.4 min step with 95%A+5% B. Then according to analytical trace a 3.6 min gradient is chosenin order to achieve good separation (e.g. from 5% to 50% B for earlyretaining compounds; from 35% to 80% B for middle retaining compoundsand so on)Wash: 1 minute wash step is performed at the end of the gradientRe-equilibration: A 2.1 minute re-equilibration step is carried out toprepare the system for the next runMake Up flow rate: 1 ml/min

Solvent:

All compounds were usually dissolved in 100% MeOH or 100% DMSO

MS Running Conditions:

Capillary voltage: 3.2 kVCone voltage: 25 V

Source Temperature: 120° C. Multiplier: 500 V Scan Range: 125-800 amuIonisation Mode ElectroSpray Positive

The starting materials for each of the Examples are commerciallyavailable unless otherwise specified.

Example 1 4-Amino-1H-pyrazole-3-carboxylic acid phenylamide 1A.4-Nitro-1H-pyrazole-3-carboxylic acid phenylamide

4-Nitropyrazole-3-carboxylic acid (2.5 g; 15.9 mmol) was added to astirred solution of aniline (1.6 ml; 17.5 mmol), EDC (3.7 g; 19.1 mmol),and HOBt (2.6 g; 19.1 mmol) in N,N-dimethylformamide (DMF) (25 ml), thenstirred at room temperature overnight. The solvent was removed byevaporation under reduced pressure and the residue triturated with ethylacetate/saturated NaHCO₃ solution. The resultant solid was collected byfiltration, washed with water and diethyl ether then dried under vacuumto give 2.85 g of the title compound (sodium salt) as a yellow/brownsolid. (LC/MS: R_(t) 2.78, [M+H]⁺ 232.95).

1B. 4-Amino-1H-pyrazole-3-carboxylic acid phenylamide

4-Nitro-1H-pyrazole-3-carboxylic acid phenylamide (100 mg; 0.43 mmol)was dissolved in ethanol (5 ml), treated with tin (II) chloridedihydrate (500 mg; 2.15 mmol) then heated at reflux overnight. Thereaction mixture was cooled and evaporated. The residue was partitionedbetween ethyl acetate and brine, and the ethyl acetate layer wasseparated, dried (MgSO₄), filtered and evaporated. The crude product waspurified by flash column chromatography eluting with 1:1 ethylacetate/petroleum ether then 5% methanol/dichloromethane. Evaporation ofproduct containing fractions followed by preparative LC/MS gave 15 mg ofthe product as an off white solid. (LC/MS: R_(t) 1.40, [M+H]⁺ 202.95).

Example 2 4-Acetylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide 2A. 4-Nitro-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

4-Nitropyrazole-3-carboxylic acid (10 g; 63.66 mmol) was added to astirred solution of 4-fluoroaniline (6.7 ml; 70 mmol), EDC (14.6 g; 76.4mmol), and HOBt (10.3 g; 76.4 mmol) in DMF (25 ml), then stirred at roomtemperature overnight. The solvent was removed by evaporation underreduced pressure and the residue triturated with ethyl acetate/saturatedbrine solution. The resultant yellow solid was collected by filtration,washed with 2M hydrochloric acid, then dried under vacuum to give 15.5 gof the title compound. (LC/MS: R_(t) 2.92 [M+H]⁺ 250.89).

2B. 4-Amino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide

4-Nitro-1H-pyrazole-3-carboxylic acid (4-fluorophenyl)-amide (15 g) wasdissolved in 200 ml of ethanol, treated with 1.5 g of 10% palladium oncarbon under a nitrogen atmosphere, then hydrogenated at roomtemperature and pressure overnight. The catalyst was removed byfiltration through Celite and the filtrate evaporated. The crude productwas dissolved in acetone/water (100 ml:100 ml) and after slowevaporation of the acetone the product was collected by filtration as abrown crystalline solid (8.1 g). (LC/MS: R_(t) 1.58, [M+H]⁺ 220.95).

2C. 4-Acetylamino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide

4-Amino-1H-pyrazole-3-carboxylic acid (4-fluorophenyl)-amide (500 mg;2.27 mmol) was dissolved in 5 ml of pyridine, treated with aceticanhydride (240 μl, 2.5 mmol) then stirred at room temperature overnight.The solvent was removed by evaporation then dichloromethane (20 ml) and2M hydrochloric acid (20 ml) were added. The undissolved solid wascollected by filtration, washed with more dichloromethane and water thendried under vacuum. The product was isolated as an off white solid (275mg). (LC/MS: R_(t) 2.96, [M+H]⁺ 262.91).

Example 3 4-(2,2,2-Trifluoro-acetylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

4-Amino-1H-pyrazole-3-carboxylic acid (4-fluorophenyl)-amide (Example2B) (500 mg; 2.27 mmol) was dissolved in 5 ml of pyridine, treated withtrifluoroacetic anhydride (320 μl, 2.5 mmol) then stirred at roomtemperature overnight. The solvent was removed by evaporation, theresidue was partitioned between ethyl acetate (50 ml) and 2 Mhydrochloric acid (50 ml), and the ethyl acetate layer was separated,washed with brine (50 ml), dried (MgSO₄), filtered and evaporated togive 560 mg of product as a brown solid. (LC/MS: [M+H]⁺ 317).

Example 44-[(5-Oxo-pyrrolidine-2-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

To a stirred solution of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluorophenyl)-amide (Example 2B) (50 mg; 0.23 mmol), EDAC (52 mg;0.27 mmol) and HOBt (37 mg; 0.27 mmol) in 5 ml of DMF was added2-oxoproline (33 mg; 0.25 mmol), and the mixture was then left at roomtemperature overnight. The reaction mixture was evaporated and theresidue purified by preparative LC/MS, to give 24 mg of the product as awhite solid. (LC/MS: R_(t) 2.27 [M+H]⁺ 332).

Example 5 4-Phenylacetylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4 butusing phenylacetic acid (34 mg; 0.23 mmol) as the starting material. Thetitle compound (14 mg) was isolated as a white solid. (LC/MS: R_(t) 3.24[M+H]⁺ 339).

Example 6 4-(2-1H-Indol-3-yl-acetylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing indole-3-acetic acid (44 mg; 0.23 mmol) as the starting material.The title product (14 mg) was isolated as a white solid. (LC/MS: R_(t)3.05 [M+H]⁺ 378).

Example 7 4-(2-Benzenesulphonyl-acetylamino)-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 2-(phenylsulphonyl)acetic acid (50 mg; 0.23 mmol) as the startingmaterial. The title compound (29 mg) was isolated as a white solid.(LC/MS: R_(t) 3.00 [M+H]⁺ 403).

Example 84-[2-(5-Amino-tetrazol-1-yl)-acetylamino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, but5-aminotetrazole-1-acetic acid (36 mg; 0.23 mmol) was used as thestarting material. The title compound (23 mg) was isolated as a whitesolid. (LC/MS: R_(t) 2.37 [M+H]⁺ 346).

Example 9N-[3-(4-Fluoro-phenylcarbamoyl)-1H-pyrazol-4-yl]-6-hydroxy-nicotinamide

The reaction was carried out in a manner analogous to Example 4, butusing 6-hydroxynicotinic acid (38 mg; 0.23 mmol) as the startingmaterial. The title compound (17 mg) was isolated as a white solid.(LC/MS: R_(t) 2.32 [M+H]⁺ 342).

Example 104-[3-(4-Chloro-phenyl)-propionylamino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 3-(4-chlorophenyl)propionic acid (46 mg; 0.23 mmol) as thestarting material. The title compound (40 mg) was isolated as a whitesolid. (LC/MS: R_(t) 3.60 [M+H]⁺ 388).

Example 114-(3-4H-[1,2,4]Triazol-3-yl-propionylamino)-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 3-triazol-3-yl propionic acid (36 mg; 0.23 mmol) as the startingmaterial. The title compound (18 mg) was isolated as a white solid.(LC/MS: R_(t) 2.39 [M+H]⁺ 344).

Example 124-[2-(1-Methyl-1H-indol-3-yl)-acetylamino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing N-methyl indole-3-acetic acid (48 mg; 0.23 mmol) as the startingmaterial. The title compound (20 mg) was isolated as a white solid.(LC/MS: R_(t) 3.34 [M+H]⁺ 392).

Example 134-[(1-Hydroxy-cyclopropanecarbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 1-hydroxycyclopropane carboxylic acid (26 mg; 0.23 mmol) as thestarting material. The title compound (24 mg) was isolated as a whitesolid. (LC/MS: R_(t) 2.55 [M+H]⁺ 305).

Example 14 1-Acetyl-piperidine-4-carboxylic acid[3-(4-fluoro-phenylcarbamoyl)-1H-pyrazol-4-yl]-amide

The reaction was carried out in a manner analogous to Example 4, butusing N-acetylpiperidine acetic acid (43 mg; 0.23 mmol) as the startingmaterial. The title compound (19 mg) was isolated as a white solid.(LC/MS: R_(t) 2.49 [M+H]⁺ 374).

Example 154-[3-(4-Methyl-piperazin-1-yl)-propionylamino]-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 4-N-methylpiperazine-1-N-propionic acid (31 mg; 0.23 mmol) as thestarting material. The title compound (19 mg) was isolated as a whitesolid. (LC/MS: R_(t) 1.77 [M+H]⁺ 375).

Example 16 4-(2-1H-Imidazol-4-yl-acetylamino)-1H-pyrazole-3-carboxylicacid (4-fluorophenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing imidazole-4-acetic acid (32 mg; 0.23 mmol) as the startingmaterial. The title compound (35 mg) was isolated as a white solid.(LC/MS: R_(t) 1.82 [M+H]⁺ 329).

Example 17 4-(3-Morpholin-4-yl-propionylamino)-1H-pyrazole-3-carboxylicacid (4-fluorophenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 3-morpholin-4-yl-propionic acid (40 mg; 0.23 mmol) as the startingmaterial. The title compound (15 mg) was isolated as a white solid.(LC/MS: R_(t) 1.84 [M+H]⁺ 362).

Example 18 4-(3-Piperidin-1-yl-propionylamino)-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide

The reaction was carried out in a manner analogous to Example 4, butusing 3-piperidine-4-yl-propionic acid (39 mg; 0.23 mmol) as thestarting material. The title compound (19 mg) was isolated as a whitesolid. (LC/MS: R_(t) 1.92 [M+H]⁺ 360).

Example 19 4-Cyclohexylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

To a solution of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide (200 mg; 1 mmol) and cyclohexanone (107 mg; 1.1mmol) in dichloromethane (10 ml) were added 3 Å molecular sieves (1 g)and sodium triacetoxyborohydride (315 mg; 1.5 mmol), and the mixture wasthen stirred at room temperature over the weekend. The reaction mixturewas filtered through Celite®, diluted with ethyl acetate, washed withbrine, dried (MgSO₄) and evaporated to give the 48 mg of the product asa grey gum. (LC/MS: R_(t) 2.95, [M+H]⁺ 285).

Example 20 4-Isopropylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The title compound was prepared in a manner analogous to Example 19, butusing acetone in place of cyclohexanone. (LC/MS: R_(t) 2.08, [M+H]⁺245).

Example 21 4-(2-Hydroxy-1-methyl-ethylamino)-1H-pyrazole-3-carboxylicacid (4-fluorophenyl)-amide

The compound was prepared in a manner analogous to Example 19, but usinghydroxyacetone in place of cyclohexanone. ¹HNMR (400 MHz, D6-DMSO): 9.9(1H, br s), 7.8 (2H, dd), 7.3 (1H, s), 7.15 (2H, t), 5.15 (1H, d), 4.7(1H, br s), 3.4 (2H, m), 3.2 (1H, m), 1.1 (3H, d).

Example 22 4-(1-Ethyl-propylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The compound was prepared in a manner analogous to Example 19, but using3-pentanone in place of cyclohexanone. ¹HNMR (400 MHz, D6-DMSO): 12.85(1 h, br s), 9.9 (1H, br s), 7.8 (2H, br t), 7.3 (1H, s), 7.15 (2H, t),5.0 (1H, d), 2.9 (1H, br m), 1.5 (4H, m), 3.2 (1H, m), 0.9 (6H, t).

Example 23 4-(3-Chloro-pyrazin-2-ylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

A mixture of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide (50 mg; 0.23 mmol) and 2,3-dichloropyrazine (140mg; 0.92 mmol) was heated at 150° C. (50 W) for 20 minutes in a CEMDiscover™ microwave synthesiser. The crude reaction mixture was purifiedby flash column chromatography eluting with ethyl acetate/hexane (1:3then 1:2). Product containing fractions were combined and evaporated togive 15 mg of the title compound as a white solid. (LC/MS: R_(t) 4.06M+H]⁺ 332).

Example 24 4-(Pyrazin-2-ylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The compound was prepared in a manner analogous to Example 23, but using2-chloropyrazine in place of 2,3-dichloropyrazine. (LC/MS: R_(t) 3.28[M+H]⁺ 299).

Example 25 Synthesis of4-(2-Methoxy-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

2-Methoxy-benzoic acid (38 mg, 0.25 mmol) was added to a solution of4-amino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide (50 mg,0.23 mmol), EDC (53 mg, 0.27 mmol), and HOBt (37 mg, 0.27 mmol) in DMF(5 ml). The reaction mixture was stirred at room temperature for 24hours. The solvent was removed under reduced pressure. The residue waspurified by preparative LC/MS and, after evaporation ofproduct-containing fractions, yielded the product as a pinkish solid (12mg, 15%). (LC/MS: R_(t) 4.00, [M+H]⁺ 354.67).

Example 26 Synthesis of 4-Benzoylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using benzoic acid (31 mg, 0.25 mmol) as starting acid. The productwas isolated as a pink solid (26 mg, 35%). (LC/MS: R_(t) 3.96, [M+H]⁺324.65).

Example 27 Synthesis of4-(Cyclohexanecarbonyl-amino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using cyclohexanecarboxylic acid (32 mg, 0.25 mmol) as starting acid.The product was isolated as a pink solid (28 mg, 37%). (LC/MS: R_(t)4.16, [M+H]⁺ 330.70).

Example 28 Synthesis of4-[(1-Methyl-cyclopropanecarbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using 1-methyl-cyclopropanecarboxylic acid (25 mg, 0.25 mmol) asstarting acid. The product was isolated as a pink solid (24 mg, 35%).(LC/MS: R_(t) 3.72, [M+H]⁺302.68).

Example 29 Synthesis of4-(2-Hydroxy-acetylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using hydroxy-acetic acid (19 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (26 mg, 41%). (LC/MS: R_(t) 2.65,[M+H]⁺ 278.61).

Example 30 Synthesis of4-(2,2-Dimethyl-propionylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using 2,2-dimethyl-propionic acid (26 mg, 0.25 mmol) as startingacid. The product was isolated as a pink solid (21 mg, 30%). (LC/MS:R_(t) 3.83, [M+H]⁺ 304.68).

Example 31 Synthesis of4-(3-Hydroxy-propionylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example25 using 3-hydroxy-propionic acid (75.1 mg, 0.25 mmol) as starting acid.The product was isolated as a beige solid (5 mg, 8%). (LC/MS: R_(t)2.58, [M+H]⁺ 292.65).

Example 32 Synthesis of4-(2-Fluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

2-Fluorobenzoic acid (36 mg, 0.25 mmol) was added to a solution of4-amino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide (50 mg,0.23 mmol), EDC (53 mg, 0.27 mmol) and HOBt (37 mg, 0.27 mmol) in DMSO(1 ml). The reaction mixture was stirred at room temperature for 24hours and purified by preparative LC/MS. Evaporation ofproduct-containing fractions yielded the product as a white solid (15mg, 19%). (LC/MS: R_(t) 3.91, [M+H]⁺ 342.66).

Example 33 Synthesis of4-(3-Fluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 3-fluorobenzoic acid (36 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (19 mg, 24%). (LC/MS: R_(t) 4.03,[M+H]⁺ 342.67).

Example 34 Synthesis of4-(3-Methoxy-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 3-methoxy-benzoic acid (39 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (20 mg, 25%). (LC/MS: R_(t) 3.97,[M+H]⁺ 354.68).

Example 35 Synthesis of4-(2-Nitro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 2-nitrobenzoic acid (43 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (17 mg, 20%). (LC/MS: R_(t) 3.67,[M+H]⁺ 369.66).

Example 36 Synthesis of4-(4-Nitro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 4-nitrobenzoic acid (43 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (15 mg, 18%). (LC/MS: R_(t) 3.98,[M+H]⁺ 369.63).

Example 37 Synthesis of4-[(3-Methyl-furan-2-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 3-methyl-2-furoic acid (32 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a white solid (15 mg, 20%). (LC/MS: R_(t) 3.86,[M+H]⁺ 328.68).

Example 38 Synthesis of4-[(Furan-2-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 2-furoic acid (29 mg, 0.25 mmol) as starting acid. The productwas isolated as a white solid (18 mg, 25%). (LC/MS: R_(t) 3.56, [M+H]⁺314.64).

Example 39 Synthesis of4-[(3H-Imidazole-4-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 1H-imidazole-4-carboxylic acid (29 mg, 0.25 mmol) as startingacid. The product was isolated as a white solid (16 mg, 22%). (LC/MS:R_(t) 2.59, [M+H]⁺ 314.65).

Example 40 Synthesis of4-(4-Fluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 4-fluorobenzoic acid (36 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a cream coloured solid (23 mg, 29%). (LC/MS:R_(t) 4.00, [M+H]⁺ 342.67).

Example 41 Synthesis of4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 2,6-difluorobenzoic acid (40 mg, 0.25 mmol) as starting acid.The product was isolated as a cream coloured solid (25 mg, 30%). (LC/MS:R_(t) 3.76, [M+H]⁺ 360.66).

Example 42 Synthesis of4-(3-Nitro-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 3-nitrobenzoic acid (43 mg, 0.25 mmol) as starting acid. Theproduct was isolated as a cream coloured solid (15 mg, 18%). (LC/MS:R_(t) 3.94, [M+H]⁺ 369.65).

Example 43 Synthesis of 1H-Indole-3-carboxylic acid[3-(4-fluoro-phenylcarbamoyl)-1H-pyrazol-4-yl]-amide

The experiment was carried out in a manner analogous to that of Example32 using indole-3-carboxylic acid (41 mg, 0.25 mmol) as starting acid.The product was isolated as a rust coloured solid (14 mg, 17%). (LC/MS:R_(t) 3.60, [M+H]⁺ 363.66).

Example 44 Synthesis of4-(4-Hydroxymethyl-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 4-hydroxymethylbenzoic acid (39 mg, 0.25 mmol) as startingacid. The product was isolated as a white solid (19 mg, 23%). (LC/MS:R_(t) 3.12, [M+H]⁺ 354.68).

Example 45 Synthesis of4-(3-Methyl-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 3-methylbenzoic acid (35 mg, 0.25 mmol) as starting acid. Theproduct was isolated as an off-white solid (21 mg, 27%). (LC/MS: R_(t)4.13, [M+H]⁺ 338.71).

Example 46 Synthesis of4-(2-Methyl-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 2-methylbenzoic acid (35 mg, 0.25 mmol) as starting acid. Theproduct was isolated as an off-white solid (20 mg, 26%). (LC/MS: R_(t)4.05, [M+H]⁺ 338.69).

Example 47 Synthesis of4-(4-Methyl-benzoylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example32 using 4-methylbenzoic acid (35 mg, 0.25 mmol) as starting acid. Theproduct was isolated as an off-white solid (19 mg, 24%). (LC/MS: R_(t)4.16, [M+H]⁺ 338.70).

Example 48 Synthesis of4-[(2-Methyl-thiophene-3-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

2-Methyl-3-thiophenecarboxylic acid (36 mg, 0.25 mmol) was added to asolution of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide (Example 2B) (50 mg, 0.23 mmol), EDC (53 mg,0.27 mmol), and HOBt (37 mg, 0.27 mmol) in DMSO (1 ml). The reactionmixture was stirred at room temperature for 24 hours. The reactionmixture was added dropwise to water (30 ml) and the resultant solid wascollected by filtration, washed with water and sucked dry. The titlecompound was obtained as a beige solid (15 mg, 19%). (LC/MS: R_(t) 4.08,[M+H]⁺ 344.67).

Example 49 Synthesis of Quinoline-2-carboxylic acid[3-(4-fluoro-phenylcarbamoyl)-1H-pyrazol-4-yl]-amide

The experiment was carried out in a manner analogous to that of Example48 using quinaldic acid (44 mg, 0.25 mmol) as starting acid. The productwas isolated as a brown solid (16 mg, 19%). (LC/MS: R_(t) 4.29, [M+H]⁺375.66).

Example 50 Synthesis of4-[(Thiophene-3-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The experiment was carried out in a manner analogous to that of Example48 using thiophene-3-carboxylic acid (33 mg, 0.25 mmol) as startingacid. The product was isolated as a beige solid (15 mg, 20%). (LC/MS:R_(t) 3.77, [M+H]⁺ 330.61).

Example 51 4-(2-fluoro-3-methoxy-benzoylamino)-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide

2-Fluoro-3-methoxybenzoic acid (0.047 g, 0.28 mmol),4-amino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide (Example2B) (0.055 g, 0.25 mmol), EDC (0.58 g, 0.30 mmol) and HOBt (0.041 g,0.30 mmol) were stirred at room temperature in DMSO (1.25 ml) for 5hours. The reaction mixture was poured into water (30 ml) and theresultant solid was collected by filtration and dried in a vacuum ovento give the title compound as a grey solid (0.058 g, 63%). (LC/MS: R_(t)3.99, [MH]⁺ 372.98).

Example 52 Synthesis of4-[2-(2-Pyrrolidin-1-yl-ethoxy)-benzoylamino]-1H-pyrazole-3-carboxylicacid 4-fluorophenylamide 52A 2-(2-Pyrrolidin-1-yl-ethoxy)-benzoic acidmethyl ester

Diisopropylazodicarboxylate (0.404 g, 2 mmol) was added dropwise to asolution of triphenylphosphine (0.524 g, 2 mmol) in THF (10 ml). Methylsalicylate (0.304 g, 2 mmol) was added dropwise and the resultantmixture was stirred at room temperature for 1 hour. 1,2-Hydroxyethylpyrrolidine (0.230 g, 2 mmol) was added dropwise and the reactionmixture was left stirring at room temperature for a further 1.5 hours.The resulting solution was reduced in vacuo and subject to flash columnchromatography, eluting with hexane:ethyl acetate (5:1, 1:1) then ethylacetate: methanol (4:1) to give the product as a clear yellow oil (0.104g, 21%). (LC/MS: R_(t) 0.69, 1.62, [MH]⁺ 250.02).

52B.4-[2-(2-Pyrrolidin-1-yl-ethoxy)-benzoylamino]-1H-pyrazole-3-carboxylicacid 4-fluorophenylamide

2-(2-Pyrrolidin-1-yl-ethoxy)-benzoic acid methyl ester (0.104 g, 0.42mmol) was treated with 2 M aqueous NaOH (20 ml) and water (20 ml). Thereaction mixture was stirred at room temperature for 20 hours, thenreduced in vacuo and azeotroped with toluene (3×5 ml). Water (50 ml) wasadded and the mixture taken to pH 5 using 1M aqueous HCl. The resultingsolution was reduced in vacuo and azeotroped with toluene (3×5 ml) togive a white solid, which was combined with4-amino-1H-pyrazole-3-carboxylic acid (4-fluoro-phenyl)-amide (Example2B) (0.055 g, 0.25 mmol), EDC (0.058 g, 0.3 mmol) and HOBt (0.041 g, 0.3mmol) and stirred at room temperature in DMSO (3 ml) for 20 hours. Thereaction mixture was poured into water (30 ml) and the resultant solidwas collected by filtration and dried in a vacuum oven to give the titlecompound as a grey solid (0.015 g, 14%). (LC/MS: R_(t) 2.18, [MH]⁺438.06).

Example 53 Synthesis of4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(1-methyl-piperidin-4-yl)-amide

A mixture of 4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(134 mg, 0.50 mmol), 4-amino-N-methylpiperidine (50.0 μl, 0.45 mmol),EDAC (104 mg, 0.54 mmol) and HOBt (73.0 mg, 0.54 mmol) in DMF (3 ml) wasstirred at ambient temperature for 16 hours. The mixture was reduced invacuo, the residue taken up in EtOAc and washed successively withsaturated aqueous sodium bicarbonate, water and brine. The organicportion was dried (MgSO₄) and reduced in vacuo to give4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid(1-methyl-piperidin-4-yl)-amide as a white solid (113 mg, 69%). (LC/MS:R_(t) 2.52, [M+H]⁺ 364.19).

Example 54 Synthesis of4-(Cyclohexyl-methyl-amino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

This compound was prepared in a manner analogous to the compound ofExample 19 by successive reductive alkylations using firstlycyclohexanone and then formaldehyde. (LC/MS: R_(t) 2.77 [MH]⁺ 316.71).

Example 55 4-(Pyridin-2-ylamino)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The title compound was prepared in a manner analogous to the compound ofExample 23. (LC/MS: R_(t) 2.07 [MH]⁺ 298.03).

Examples 56-81

By following the procedures described in the foregoing examples ormethods analogous thereto, or by carrying out chemical transformationsusing the compounds described in the above examples and syntheticmethods well known to the skilled person, the compounds set out in Table3 were prepared.

TABLE 3 Prepared using method Example analogous to Differences to No.Structure Example No Example? LCMS 56

 4 R_(t) 3.20 min [M + H]⁺ 406.07 57

 4 The removal of t-Boc protecting group with TFA as described inExample 82 R_(t) 2.35 min m/z 343.72 58

 4 Used DMSO instead of DMF as solvent R_(t) 3.51 min m/z 314.62 59

 4 Used DMSO instead of DMF as solvent R_(t) 3.79 min m/z 363.67 60

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 3.68 min m/z 384.69 61

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 3.61 min m/z 326.10 62

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 3.51 min m/z 387.11 63

48 R_(t) 3.11 min m/z 313.65 64

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 2.20 min m/z 455.19 65

53 R_(t) 3.95 min m/z 349.09 66

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 2.39 min m/z 351.07 67

48 Purified by column chromatography using EtOAc: Petroleum ether eluentR_(t) 2.83 min m/z 365.13 68

Removal of PMB group from the compound of Example 62 using TFA- anisoleR_(t) 2.10 min m/z 266.97 69

48 Used DMF instead of DMSO as solvent R_(t) 3.22 min m/z 363.10 70

48 R_(t) 4.48 min m/z 358.96 71

48 R_(t) 3.93 min m/z 340.96 72

48 R_(t) 4.11 min m/z 373.01 73

48 Used DMF instead of DMSO as solvent R_(t) 2.56 min m/z 373.05 74

Obtained by oxidation and then reductive amination of Example 73 R_(t)1.99 min m/z 442.09 75

53 Purified by column chromatography using DCM:MeOH (1:0 to 19:1) eluentR_(t) 3.65 min m/z 335.03 76

25 Purified by column chromatography. The removal of t-Boc protectinggroup with saturated ethyl acetate/HCl R_(t) 1.57 min m/z 350.10 77

53 R_(t) 5.05 min m/z 405.14 78

53 R_(t) 2.87 min m/z 416.07 79

53 Purified by column chromatography using EtOAc: Petroleum ether eluent(1:1) R_(t) 3.41 min m/z 321.03 80

2A, 2B & 53 Commercially available 5- methyl-pyrazole- 1H-3-carboxylicacid used as starting material. Purified by column chromatography usingEtOAc: Hexane eluent (1:3 to 1:1) R_(t) 3.42 min m/z 375.05 81

2C Purified by column chromatography using EtOAc: Hexane eluent (1:1 to1:0) R_(t) 2.37 min m/z 277.04

Example 82 4-[(4-Amino-1-methyl-1H-imidazole-2-carbonyl)-amino]-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

Trifluoroacetic acid (200 μl) was added to a stirred suspension of{2-[3-(4-fluoro-phenylcarbamoyl)-1H-pyrazol-4-ylcarbamoyl]-1-methyl-1H-imidazol-4-yl}-carbamicacid tert-butyl ester (30 mg) in dichloromethane (5 ml), then stirred atroom temperature for 2 hours. The solvent was evaporated thenre-evaporated with toluene (2×10 ml). The residue was triturated withdiethyl ether and the resultant solid collected by filtration. The solidwas washed with diethyl ether then dried under vacuum to give 15 mg of4-[(4-amino-1-methyl-1H-imidazole-2-carbonyl)-amino]-1H-pyrazole-3-carboxylicacid (4-fluoro-phenyl)-amide as an off-white solid. (LC/MS: [M+H]⁺343.72).

Example 83 Synthesis of4-{[4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carbonyl]amino}-cyclohexanecarboxylicacid 83A.4-{[4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carbonyl]amino}-cyclohexanecarboxylicacid ethyl ester

Thionyl chloride (0.32 ml, 4.40 mmol) was slowly added to a mixture of4-aminocyclohexanecarboxylic acid (572 mg, 4.00 mmol) in EtOH (10 ml)and stirred at ambient temperature for 16 hours. The mixture was reducedin vacuo, azeotroping with toluene, to give the corresponding ethylester (650 mg) as a pale solid.

A mixture of the ethyl ester (103 mg, 0.60 mmol),4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid (134 mg,0.50 mmol), EDC (115 mg, 0.60 mmol) and HOBt (81 mg, 0.60 mmol) in DMF(5 ml) was stirred at ambient temperature for 16 hours. The mixture wasreduced in vacuo, the residue taken up in EtOAc and washed successivelywith saturated aqueous sodium bicarbonate, water and brine. The organicportion was dried (MgSO₄) and reduced in vacuo to give4-{[4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-cyclohexanecarboxylicacid ethyl ester (112 mg).

83B.4-{[4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-cyclohexanecarboxylicacid

A mixture of the ester (45 mg) (from 83A) in MeOH (2.5 ml) and 2Maqueous NaOH (2.5 ml) was stirred at ambient temperature for 16 hours.The volatiles were removed in vacuo, water (10 ml) added and the mixturetaken to pH 5 using 1M aqueous HCl. The precipitate formed was collectedby filtration and purified by column chromatography using EtOAc/MeOH(1:0-9:1) to give4-{[4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-cyclohexanecarboxylicacid (11 mg) as a white solid and mixture of cis-/trans-isomers. (LC/MS:R_(t) 2.78 and 2.96, [M+H]⁺ 393.09).

Examples 84-152 General Procedure A Preparation of Amide from PyrazoleCarboxylic Acid

A mixture of the appropriate benzoylamino-1H-pyrazole-3-carboxylic acid(0.50 mmol), EDAC (104 mg, 0.54 mmol), HOBt (73.0 mg, 0.54 mmol) and thecorresponding amine (0.45 mmol) in DMF (3 ml) was stirred at ambienttemperature for 16 hours. The mixture was reduced in vacuo, the residuetaken up in EtOAc and washed successively with saturated aqueous sodiumbicarbonate, water and brine. The organic portion was dried (MgSO₄) andreduced in vacuo to give the desired product.

General Procedure B Preparation of Amide from Amino-Pyrazole

To a stirred solution of the appropriate4-amino-1H-pyrazole-3-carboxylic acid amide (0.23 mmol), EDAC (52 mg;0.27 mmol) and HOBt (37 mg; 0.27 mmol) in 5 ml of N,N-dimethylformamidewas added the corresponding carboxylic acid (0.25 mmol), and the mixturewas then left at room temperature overnight. The reaction mixture wasevaporated and the residue purified by preparative LC/MS, to give theproduct.

General Procedure C Deprotection of Piperidine Ring Nitrogen by Removalof Cert-Butoxycarbonyl Group

A product of Procedure A or Procedure B containing a piperidine groupbearing an N-tert-butoxycarbonyl (t-Boc) protecting group (40 mg) wastreated with saturated ethyl acetate/HCl, and stirred at roomtemperature for 1 hour. A solid precipitated out of the reactionmixture, which was filtered off, washed with ether, and then dried togive 25 mg product (LC/MS: [M+H]⁺ 364).

Procedure L Preparation of Amine Starting Materials

The following method was used to prepare the following amines:

-   4-thiomorpholine-4-yl-cyclohexylamine;-   4-(1,1-dioxo-thiomorpholine-4-yl)-cyclohexylamine;-   N-(tetrahydro-pyran-4-yl)-cyclohexane-1,4-diamine;-   4-(4-methyl-piperazin-1-yl)-cyclohexylamine;-   1′-methyl-[1,4′]bipiperidinyl-4-ylamine; and-   4-morpholin-4-yl-cyclohexylamine.

A solution of N-4-Boc-aminocyclohexanone (0.5 g, 2.3 mmol) in THF (10ml) was treated with the appropriate amine, e.g. thiomorpholine (0.236g, 2.3 mmol), and sodium triacetoxyborohydride (0.715 g, 2.76 mmol) andacetic acid (0.182 ml). The reaction was stirred overnight at roomtemperature, then diluted with CH₂Cl₂ and washed with saturated sodiumcarbonate. The organic layer was dried over MgSO₄ and evaporated to givea white solid which was used without further purification in the nextstep. The white solid was treated with saturated HCl/EtOAc, stirred atroom temperature for 1 hour, evaporated to dryness and thenre-evaporated with toluene. The resulting amines were isolated as thehydrochloride salt. (LC/MS: R_(t) 1.75, [M+H]⁺ 201).

By following General Procedures A, B, C and L, modified where stated,the compounds set out in Table 4 were prepared.

TABLE 4 Example No. Method of Preparation LCMS  84

Procedure A [M + H]⁺ 380 R_(t) 1.42  85

Procedure A [M + H]⁺ 426 R_(t) 1.93  86

Procedure A [M + H]⁺ 440 R_(t) 1.87  87

Procedure A [M + H]⁺ 406 R_(t) 2.78  88

Procedure A [M + H]⁺ 406 R_(t) 2.55  89

Procedure A DMSO instead of DMF [M + H]⁺ 358 R_(t) 1.98  90

Procedure A DMSO instead of DMF [M + H]⁺ 357 R_(t) 3.37  91

Procedure A DMSO instead of DMF [M + H]⁺ 391 R_(t) 3.16  92

Procedure A DMSO instead of DMF [M + H]⁺ 375 R_(t) 3.02  93

Procedure A DMSO instead of DMF [M + H]⁺ 425 R_(t) 3.27  94

Procedure A DMSO instead of DMF [M + H]⁺ 393 R_(t) 3.01  95

Procedure A DMSO instead of DMF [M + H]⁺ 365 R_(t) 2.22  96

Procedure A DMSO instead of DMF [M + H]⁺ 387 R_(t) 3.05  97

Procedure A DMSO instead of DMF [M + H]⁺ 464 R_(t) 3.17  98

Procedure C using the product of Example 97 as starting material [M +H]⁺ 364 R_(t) 1.76  99

Procedure A DMSO instead of DMF [M + H]⁺ 389 R_(t) 2.36 100

Procedure A DMSO instead of DMF [M + H]⁺ 351 R_(t) 2.55 101

Procedure A DMSO instead of DMF [M + H]⁺ 362 R_(t) 2.63 102

Procedure A DMSO instead of DMF Starting amine prepared according toProcedure L [M + H]⁺ 364 R_(t) 1.75 103

Procedure A DMSO instead of DMF [M + H]⁺ 358 R_(t)3.2 104

Procedure A DMSO instead of DMF [M + H]⁺ 358 R_(t) 1.77 105

Procedure A DMSO instead of DMF [M + H]⁺ 344 R_(t) 2.71 106

Procedure A DMSO instead of DMF [M + H]⁺ 392 R_(t) 2.57 107

Procedure A DMSO instead of DMF [M + H]⁺ 347 R_(t) 2.8 108

Procedure A DMSO instead of DMF [M + H]⁺ 371 R_(t)3.1 109

Procedure A Et₃N 1 equiv., DMSO instead of DMF [M + H]⁺ 404 R_(t) 2.7110

Procedure A Et₃N 2 equiv., HOAt instead of HOBt, DMSO instead of DMF[M + H]⁺ 428 R_(t) 2.63 111

Procedure Procedure A followed by Procedure C Et₃N 2 equiv., HOAtinstead of HOBt, DMSO instead of DMF [M + H]⁺ 364 R_(t) 1.75 112

Procedure A Et₃N 2 equiv., HOAt instead of HOBt, DMSO instead of DMF[M + H]⁺ 427 R_(t) 2.71 113

Procedure A HOAt instead of HOBt, DMSO instead of DMF [M + H]⁺ 363 R_(t)3.34 114

Procedure A Et₃N 2 equiv., HOAt instead of HOBt, DMSO instead of DMF[M + H]⁺ 432 R_(t) 2.63 115

Procedure A [M + H]⁺ 461 R_(t) 3.3 116

Procedure A DMSO instead of DMF, Et₃N 2 equiv Starting amine preparedaccording to Procedure L [M + H]⁺ 448 R_(t) 1.87 117

Procedure A DMSO instead of DMF, Et₃N 2 equiv Starting amine preparedaccording to Procedure L [M + H]⁺ 447 R_(t) 1.65 118

Procedure A DMSO instead of DMF, Et₃N 2 equiv Starting amine preparedaccording to Procedure L [M + H]⁺ 447 R_(t) 1.72 119

Procedure B [M + H]⁺ 462 R_(t) 2.97 120

Procedure A N-ethyl-morpholine (NEM) 2 equiv [M + H]⁺ 379 R_(t) 2.45 121

Procedure A HOAt instead of HOBt, Et₃N 2 equiv Starting amine preparedaccording to Procedure L [M + H]⁺ 450 R_(t) 1.97 122

Procedure B [M + H]⁺ 387 R_(t) 3.83 123

Procedure B [M + H]⁺ 417 R_(t) 3.65 124

Procedure A HOAt instead of HOBt, Et₃N 2 equiv [M + H]⁺ 392 R_(t) 1.85125

Procedure A HOAt instead of HOBt, Et₃N 2 equiv [M + H]⁺ 408 R_(t) 1.82126

Procedure B [M + H]⁺ 403 R_(t) 4.02 127

Procedure B [M + H]⁺ 369 R_(t) 3.78 128

Procedure B [M + H]⁺ 435 R_(t) 3.83 129

Procedure B [M + H]⁺ 405 R_(t) 3.96 130

Procedure A HOAt instead of HOBt [M + H]⁺ 512 R_(t)3.1 131

Procedure A HOAt instead of HOBt, [M + H]⁺ 428 R_(t)2.45 132

Procedure A HOAt instead of HOBt, Et₃N 2 equiv. Cis and trans isomersseparated after amide coupling step Starting amine prepared according toProcedure L [M + H]⁺ 482 R_(t)1.96 133

Procedure A HOAt instead of HOBt, DMSO instead of DMF [M + H]⁺ 434R_(t)2.3 134

Procedure B [M + H]⁺ 442 R_(t) 2.39 135

Procedure B [M + H]⁺ 458 R_(t) 2.26 136

Procedure B HOAt instead of HOBt, [M + H]⁺ 468 R_(t)3.07 137

Procedure A Et₃N 2 equiv., HOAt instead of HOBt, [M + H]⁺ 379 R_(t)2.6138

Procedure B [M + H]⁺ 472 R_(t) 2.40 139

Procedure A Et₃N equiv., HOAt instead of HOBt, DMSO instead of DMF [M +H]⁺ 364 R_(t)2.1 140

Procedure B followed by Procedure C [M + H]⁺ 314 R_(t) 1.78 141

Procedure B followed by Procedure C [M + H]⁺ 332 R_(t) 1.89 142

Procedure B followed by Procedure C [M + H]⁺ 362 R_(t) 1.78 143

Procedure B followed by Procedure C [M + H]⁺ 348 R_(t) 2.01 144

Procedure B followed by Procedure C [M + H]⁺ 350 R_(t) 1.97 145

Procedure B followed by Procedure C [M + H]⁺ 380 R_(t) 2.01 146

Procedure B followed by Procedure C [M + H]⁺ 395 R_(t) 1.94 147

Procedure B followed by Procedure C [M + H]⁺ 396 R_(t) 2.11 148

Procedure B followed by Procedure C HOAt instead of HOBt [M + H]⁺ 368R_(t)1.76 149

Procedure B followed by Procedure C [M + H]⁺ 366 R_(t) 1.78 150

Procedure B followed by Procedure C [M + H]⁺ 383 R_(t) 1.87 151

Procedure B followed by Procedure C [M + H]⁺ 433 R_(t) 1.89 152

Procedure B followed by Procedure C HOAt instead of HOBt [M + H]⁺ 350R_(t)1.76

Examples 153-165 General Procedure D Preparation of Protected4-Amino-pyrazol-3-yl carboxylic acid 4-hydroxy-cyclohexylamide

Step D (i)

A mixture of 4-nitro-3-pyrazolecarboxylic acid (4.98 g, 31.7 mmol),trans 4-aminocyclohexanol (3.65 g, 31.7 mmol), EDAC (6.68 g, 34.8 mmol)and HOBt (4.7 g, 34.8 mmol) in DMF (120 ml) was stirred at ambienttemperature for 16 hours. The mixture was reduced in vacuo, the residuetaken up in CH₂Cl₂ and washed successively with 5% citric acid,saturated aqueous sodium bicarbonate, water and brine. The product wasfound to be mainly in the citric acid wash, which was basified andextracted with EtOAc. The organic layer was dried over MgSO₄, filteredand evaporated to give a white solid, which was triturated with CHCl₃ togive 1.95 g of 4-nitro-1H-pyrazole-3-carboxylic acid4-hydroxy-cyclohexylamide. (LC/MS: R_(t) 1.62, [M+H]⁺ 255).

Step D (ii) Introduction of Tetrahydro-pyran-2-yl Protecting Group

A solution of 4-nitro-1H-pyrazole-3-carboxylic acid4-hydroxy-cyclohexylamide (1.95 g; 7.67 mmol) in a mix of THF (50 ml)and chloroform (100 ml), was treated with 3,4-dihydro-2H-pyran (1.54 ml,15.34 mmol) and p-toluenesulphonic acid monohydrate (100 mg). Thereaction mixture was stirred at room temperature overnight, and thenexcess pyran (0.9 ml) was added in total to bring reaction tocompletion. The reaction mixture was diluted with CH₂Cl₂ and washedsuccessively with saturated aqueous sodium bicarbonate, water and brine.The resulting solution was reduced in vacuo and subject to Biotagecolumn chromatography, eluting with hexane (2 column lengths) followedby 30% ethyl acetate:hexane (10 column lengths), 70% ethylacetate:hexane (10 column lengths) to give 1.25 g of4-nitro-1-(tetrahydro-pyran-2-yl-1H-pyrazole-3-carboxylic acid[4-(tetrahydro-pyran-2-yloxy)-cyclohexyl]-amide. (LC/MS: R_(t) 2.97,[M+H]⁺ 423).

Step D (iii)

A solution of 4-nitro-1-(tetrahydro-pyran-2-yl)-1H-pyrazole-3-carboxylicacid [4-(tetrahydro-pyran-2-yloxy)-cyclohexyl]-amide (0.3 g; 0.71 mmol)in methanol (25 ml), was treated with 10% palladium on carbon (30 mg)then hydrogenated at room temperature and pressure overnight. Thecatalyst was removed by filtration and washed three times with methanol.The filtrate was evaporated to give 0.264 g of the required product.(LC/MS: R_(t) 2.39, [M+H]⁺ 393).

General Procedure E Procedure for Removal of a Tetrahydropyran-2-ylProtecting Group

To a suspension of4-(2-methoxy-benzoylamino)-1-(tetrahydro-pyran-2-yl-1H-pyrazole-3-carboxylicacid [4-(tetrahydro-pyran-2-yloxy)-cyclohexyl]-amide (0.125 g, 0.23mmol) in EtOH (10 ml) was added p-toluene sulphonic acid hydrate (90 mg,0.46 mmol). The reaction mixture was heated at 70° C. for 30 mins. Thereaction was diluted with EtOAc and washed successively with saturatedaqueous sodium bicarbonate, water and brine. The resulting solution wasreduced in vacuo to give a white solid, which contained traces ofp-toluene sulphonic acid hydrate. The solid was then taken up in EtOAcand washed with 1M NaOH and then brine. The resulting solution wasreduced in vacuo and then triturated with ether/hexane to give 10 mg ofrequired product. (LC/MS: R_(t) 2.29, [M+H]⁺ 359)

General Procedure F Preparation of a Urea from a4-Amino-pyrazole-3-carboxylic acid amide

To a solution of4-amino-1-(tetrahydro-pyran-2-yl-1H-pyrazole-3-carboxylic acid[4-(tetrahydro-pyran-2-yloxy)-cyclohexyl]-amide (80 mg, 0.2 mmol) intoluene (2 ml) was added phenyl isocyanate (929 mg, 0.24 mmol). Thereaction mixture was heated at 70° C. for 1 hour. The reaction wasdiluted with EtOAc and washed successively with water and brine. Theresulting solution was reduced in vacuo to give yellow oil. This wasused without further purification. (LC/MS: R_(t) 2.28, [M+H]⁺ 344).

General Procedure G Conversion of a 4-Amino-pyrazole group to a4-(Morpholine-4-carbonylamino)-Pyrazole Group

To a solution of4-amino-1-(tetrahydro-pyran-2-yl-1H-pyrazole-3-carboxylic acid[4-(tetrahydro-pyran-2-yloxy)-cyclohexyl]-amide (0.1 g, 0.255 mmol) inCH₂Cl₂ (5 ml) at −10° C. was added in a dropwise manner a 20% solutionof phosgene in toluene. The reaction mixture was stirred at −10° C. for15 mins and then morpholine (0.765 mmol) was added. The reaction mixturewas allowed to warm up to room temperature over 1 hour then stirred atroom temperature overnight. The reaction was diluted with CH₂Cl₂ andwashed successively with saturated sodium bicarbonate and brine. Theresulting solution was reduced in vacuo to give a yellow oil which wasused without further purification. (LC/MS: R_(t) 1.68, [M+H]⁺ 338).

General Procedure H Preparation of N-Oxides

To a suspension of the compound of Example 53 (7.7 mg, 0.02 mmol) inCH₂Cl₂ (0.5 ml) was added meta-chloroperbenzoic acid (MCPBA) (3.6 mg,0.02 mmol). The reaction mixture was stirred at room temperatureovernight, and then evaporated. The residue was purified by preparativeLC/MS, to give 3 mg of the required product. (LC/MS: R_(t) 1.83, [M+H]⁺380)

General Procedure I Removal of a Benzyloxycarbonyl Protecting Group

A solution of the compound of Example 130 (0.2 g; 0.39 mmol) in EtOAc(40 ml) was treated with 10% palladium on carbon (20 mg) thenhydrogenated at room temperature and pressure for 3 hours. The catalystwas removed by filtration and washed three times with EtOAc. Thefiltrate was evaporated and the residue was subjected to chromatographyusing 10% MeOH—CH₂Cl₂ then 20% MeOH—CH₂Cl₂ to give 80 mg of the requiredproduct. (LC/MS: R_(t) 1.88, [M+H]⁺ 378).

General Procedure J Mesylation of an Amine

To a solution of the compound of Example 163 (20 mg, 0.05 mmol) in CH₃CN(3 ml) added methane-sulphonyl chloride (0.0045 ml, 0.058 mmol) followedby Hunig's Base (0.018 ml, 0.1 mmol). The reaction mixture was stirredat room temperature for 2 hours and was then evaporated down. Theresidue was purified by preparative LC/MS to give 8 mg of the requiredproduct. (LC/MS: R_(t) 2.54, [M+H]⁺ 456).

By following Procedures A to L, the compounds set out in Table 5 wereprepared.

TABLE 5 Example No. Method of Preparation LCMS 153

Procedure D followed by B then E HOAt instead of HOBt, CH₂Cl₂ instead ofDMF [M + H]⁺ 359 R_(t) 2.29 154

Procedure D followed by B then E HOAt instead of HOBt, CH₂Cl₂ instead ofDMF [M + H]⁺ 377 R_(t) 2.22 155

Procedure D followed by B then E HOAt instead of HOBt, CH₂Cl₂ instead ofDMF [M + H]⁺ 381 R_(t) 2.34 156

Procedure D followed by F then E [M + H]⁺ 344 R_(t) 2.28 157

Procedure D followed by F then E [M + H]⁺ 358 R_(t) 2.22 158

Procedure D followed by B then E HOAt instead of HOBt, CH₂Cl₂ instead ofDMF [M + H]⁺ 365 R_(t) 2.21 159

Procedure D followed by B then E HOAt instead of HOBt, CH₂Cl₂ instead ofDMF [M + H]⁺ 387 R_(t) 2.29 160

Procedure D followed by F then E [M + H]⁺ 380 R_(t) 2.17 161

Procedure D followed by G then E [M + H]⁺ 338 R_(t) 1.68 162

Procedure H [M + H]⁺ 380 R_(t) 1.83 163

Procedure A (HOAt instead of HOBt) to give the compound of Example 130followed by Procedure 1. [M + H]⁺ 378 R_(t) 1.78 164

Procedures A (HOAt instead of HOBt) and I to give the compound ofExample 163 followed by Procedure J [M + H]⁺ 456 R_(t) 2.54

General Procedure M Formation of Pyrazole 4-Amide Group

4-Nitropyrazole-3-carboxylic acid (7.3 g; 15.9 mmol) was added to astirred solution of 4-amino-1-Boc-piperidine (10.2 mg; 51 mmol), EDC(10.7 g; 55.8 mmol), and HOAt (55.8 g; 19.1 mmol) in DMF (100 ml), andthen stirred at room temperature overnight. The solvent was removed byevaporation under reduced pressure and the residue triturated with water(250 ml). The resultant cream solid was collected by filtration, washedwith water then dried under vacuum to give 13.05 g of4-[(4-nitro-1H-pyrazole-3-carbonyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester (LC/MS: R_(t) 2.50, [M+H]⁺ 340).

4-[(4-Nitro-1H-pyrazole-3-carbonyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester (13.05 g) was dissolved in ethanol/DMF (300 ml/75 ml),treated with 10% palladium on carbon (500 mg) then hydrogenated at roomtemperature and pressure overnight. The catalyst was removed byfiltration through Celite and the filtrate evaporated and re-evaporatedwith toluene. The crude material was purified by flash columnchromatography eluting with EtOAc then 2% MeOH/EtOAc then 5% MeOH/EtOAc.Product containing fractions were combined and evaporated to give 8.78 gof 4-[(4-amino-1H-pyrazole-3-carbonyl)-amino]-piperidine-1-carboxylicacid tert-butyl ester as a brown foam. (LC/MS: R_(t) 1.91, [M+H]⁺ 310).

To a stirred solution of4-[(4-amino-1H-pyrazole-3-carbonyl)-amino]-piperidine-1-carboxylic acidtert-butyl ester (200 mg; 0.65 mmol), EDAC (150 mg; 0.78 mmol) and HOBt(105 mg; 0.78 mmol) in 5 ml of N,N-dimethylformamide was added thecorresponding carboxylic acid (0.25 mmol), and the mixture was then leftat room temperature overnight. The reaction mixture was diluted withsaturated aqueous sodium bicarbonate solution and the product collectedby filtration and dried under vacuum. The Boc-protected compound wasdissolved in saturated HCl/EtOAc and stirred at room temperature for 3hours. The product was collected by filtration, washed with diethylether and dried under vacuum.

General Procedure N Preparation of 1-tert-Butyl-piperidin-4-ylamine

Step N (i)

To a solution of 1-ethyl-4-oxopiperidine (25 g, 0.197 mol) in acetone(250 ml) at RT in a water bath was added methyl iodide (15.5 ml, 0.25mol) at such a rate to keep the temperature below 30° C. The mixture wasfiltered and the precipitate washed with acetone and dried to yield1-ethyl-1-methyl-4-oxopiperidinium iodide (45 g) (LC/MS: R_(t) 0.38,[M+H]⁺ 143).

Step N (ii)

To a solution of t-butylamine (78.2 ml, 0.74 mol) in toluene (400 ml)was added a solution of 1-ethyl-1-methyl-4-oxopiperidinium iodide (40 g,0.148 mol) and sodium bicarbonate (1.245 g, 0.014 mol) in water (60 ml).The reaction mixture was heated at 78° C. for 6 hours and then allowedto cool to ambient temperature. The layers were separated and theaqueous layer was washed with EtOAc. The organics were combined andwashed with brine, dried (MgSO₄), filtered and reduced in vacuo to yield1-tert-butyl-4-oxopiperidine (14 g) (LC/MS: R_(t) 0.39, [M+H]⁺ 156).

Step N (iii)

A solution of 1-tert-butyl-4-oxopiperidine (3.6 g, 23.1), benzylamine(5.1 ml, 46.8 mmol), acetic acid (1.5 ml) and sodiumtriacetoxyborohydride (7.38 g, 34.8 mmol) was stirred at ambient for 2days. Reaction mixture reduced in vacuo, residue partitioned betweenaqueous K₂CO₃ and EtOAc. The organic portion was dried (Na₂SO₄),filtered and reduced in vacuo. The residue was subjected tochromatography using CH₂Cl₂/MeOH/NH₄OH (87/12/1) as the eluent to yieldN-benzyl-1-tert-butylpiperidin-4-amine (1.5 g) (LC/MS: R_(t) 0.45,[M+H]⁺ 247).

Step N (iv)

A solution of N-benzyl-1-tert-butylpiperidin-4-amine (1.56 g) and 10%palladium on carbon (2 g) in MeOH (250 ml) was hydrogenated in a Parrshaker at 50 psi for 16 hours. The solution was filtered and thereaction mixture reduced in vacuo, to yield1-tert-butylpiperidin-4-amine (0.64 g) (LC/MS: R_(t) 02.31, [M+H]⁺).

Example 165 Synthesis of4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenyl]-amide 165A. Synthesisof 4-nitro-1H-pyrazole-3-carboxylic acid ethyl ester

Thionyl chloride (2.90 ml, 39.8 mmol) was slowly added to a mixture of4-nitro-3-pyrazolecarboxylic acid (5.68 g, 36.2 mmol) in EtOH (100 ml)at ambient temperature and the mixture stirred for 48 h. The mixture wasreduced in vacuo and dried through azeotrope with toluene to afford4-nitro-1H-pyrazole-3-carboxylic acid ethyl ester as a white solid (6.42g, 96%). (¹H NMR (400 MHz, DMSO-d₆) δ 14.4 (s, 1H), 9.0 (s, 1H), 4.4 (q,2H), 1.3 (t, 3H)).

165B. Synthesis of 4-amino-1H-pyrazole-3-carboxylic acid ethyl ester

A mixture of 4-nitro-1H-pyrazole-3-carboxylic acid ethyl ester (6.40 g,34.6 mmol) and 10% Pd/C (650 mg) in EtOH (150 ml) was stirred under anatmosphere of hydrogen for 20 h. The mixture was filtered through a plugof Celite, reduced in vacuo and dried through azeotrope with toluene toafford 4-amino-1H-pyrazole-3-carboxylic acid ethyl ester as a pink solid(5.28 g, 98%). (¹H NMR (400 MHz, DMSO-d₆) δ 12.7 (s, 1H), 7.1 (s, 1H),4.8 (s, 2H), 4.3 (q, 2H), 1.3 (t, 3H)).

165C. Synthesis of4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid ethyl ester

A mixture of 2,6-difluorobenzoic acid (6.32 g, 40.0 mmol),4-amino-1H-pyrazole-3-carboxylic acid ethyl ester (5.96 g, 38.4 mmol),EDC (8.83 g, 46.1 mmol) and HOBt (6.23 g, 46.1 mmol) in DMF (100 ml) wasstirred at ambient temperature for 6 h. The mixture was reduced invacuo, water added and the solid formed collected by filtration andair-dried to give 4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylicacid ethyl ester as the major component of a mixture (15.3 g). (LC/MS:R_(t) 3.11, [M+H]⁺ 295.99).

165D. Synthesis of4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid

A mixture of 4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acidethyl ester (10.2 g) in 2 M aqueous NaOH/MeOH (1:1, 250 ml) was stirredat ambient temperature for 14 h. Volatile materials were removed invacuo, water (300 ml) added and the mixture taken to pH 5 using 1Maqueous HCl. The resultant precipitate was collected by filtration anddried through azeotrope with toluene to afford4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid as a pinksolid (5.70 g). (LC/MS: R_(t) 2.33, [M+H]⁺ 267.96).

165E. Synthesis of 5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenylamine

3,4-Dinitrofluorobenzene (1.86 g, 10 mmol) and4-hydroxy-1-methylpiperidine (1.38 g, 12 mmol) were dissolved in THF (20ml) and stirred at ambient temperature while sodium hydride (60%dispersion in mineral oil, 0.40 g, 10 mmol) was added in several smallportions. The reaction mixture was stirred for one hour and then reducedin vacuo, partitioned between ethyl acetate and water, and the organicphase washed with brine, dried (MgSO₄) and reduced in vacuo. Theresulting residue was subject to column chromatography, eluting with 5%MeOH/DCM to give a yellow solid (1.76 g, 2:1 ratio of4-(3,4-dinitro-phenoxy)-1-methyl-piperidine and a4-(4-fluoro-2-nitro-phenoxy)-1-methyl-piperidine).

A sample of the mixture of products obtained (0.562 g) was dissolved inDMF (10 ml) under an atmosphere of nitrogen. Palladium on carbon (10%,0.056 g) was added and the reaction mixture was shaken under a hydrogenatmosphere for 40 hours. The solids were removed by filtration and thefiltrate reduced in vacuo, taken up in ethyl acetate, washed (saturatedaqueous ammonium chloride solution, then saturated aqueous brine), dried(MgSO₄) and reduced in vacuo to give5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenylamine) as a brown oil(0.049 g, 7%). (¹H NMR (400 MHz, MeOD-d₄) δ 6.6 (m, 2H), 6.4 (m, 1H),4.3 (m, 1H), 2.7 (m, 2H), 2.3 (m, 2H), 1.9 (m, 2H), 1.7 (m, 2H)).

165F. Synthesis of4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenyl]-amide

5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenylamine) (0.049 g, 0.22mmol) was combined with4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid (0.053 g,0.20 mmol), EDC (0.048 g, 0.25 mmol), HOBt (0.034 g, 0.25 mmol) and DMF(1 ml) and the resulting reaction mixture was stirred at ambienttemperature for 18 hours. The reaction mixture was reduced in vacuo andpurified by preparative LC/MS to give4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[5-fluoro-2-(1-methyl-piperidin-4-yloxy)-phenyl]-amide as a buff solid.(0.010 g, 11%) (LC/MS: R_(t) 2.19, [M+H]⁺ 474.27).

Example 166 Synthesis of4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[5-fluoro-2-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-amide

3,4-Dinitrofluorobenzene (0.93 g, 5 mmol) and1-(2-hydroxyethylpyrrolidine) (0.69 g, 6 mmol) were dissolved in THF (10ml) and stirred at ambient temperature while sodium hydride (60%dispersion in mineral oil, 0.24 g, 6 mmol) was added in several smallportions. The reaction mixture was stirred for 5 hours, diluted withethyl acetate and the combined organics washed with water and brine,dried (MgSO₄) and reduced in vacuo. The resulting residue was subject tocolumn chromatography, eluting with 5% MeOH/DCM to give an orange oil(0.94 g, 1:1 ratio of 1-[2-(3,4-dinitro-phenoxy)-ethyl]-pyrrolidine and1-[2-(4-Fluoro-2-nitro-phenoxy)-ethyl]-pyrrolidine.

A sample of the mixture of products obtained (0.281 g) was dissolved inDMF (5 ml) under an atmosphere of nitrogen. Palladium on carbon (10%,0.028 g) was added and the reaction mixture was shaken under a hydrogenatmosphere for 20 hours. The solids were removed by filtration and thefiltrate reduced in vacuo and combined with4-(2,6-difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid (0.134 g,0.50 mmol), EDC (0.116 g, 0.60 mmol), HOBt (0.081 g, 0.60 mmol) and DMF(2.5 ml) and the resulting reaction mixture was stirred at ambienttemperature for 18 hours. The reaction mixture was reduced in vacuo andthe residue partitioned between ethyl acetate (50 ml) and saturatedaqueous sodium bicarbonate solution (50 ml). The organic layer waswashed with brine, dried (MgSO₄) and reduced in vacuo to give theintermediate amides. Acetic acid (10 ml) was added to the crude amideand the mixture was heated at reflux for 3 hours and then reduced invacuo. 4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[5-fluoro-2-(2-pyrrolidin-1-yl-ethoxy)-phenyl]-amide was isolated fromthe residue by preparative LC/MS as an off white solid (0.040 g, 5.6%).(LC/MS: R_(t) 2.38, [M+H]⁺ 474.33).

Examples 167-223

By following the procedures described above, the compounds set out inTable 6 were prepared.

TABLE 6 Example No. Structure Method Differences LCMS 167

A Starting amine prepared according to Procedure L HOAt instead of HOBtDMSO as solvent instead of DMF Et₃N 2 eq Purified by HPLC Cis/TransIsomers separated after amine preparation (L) [M + H]⁺ 434 R_(t) 1.97168

A Starting amine prepared according to Procedure L HOAt instead of HOBtDMSO as solvent instead of DMF Et₃N 2 eq Purified by chromatography 10%MeOH/ CH₂Cl₂ Cis/Trans Isomers separated after amine preparation (L)[M + H]⁺ 434 R_(t) 2.03 169

Procedure D followed by G then E [M + H]⁺ 338 R_(t) 2.28 170

A Starting amine prepared according to Procedure L DMSO as solventinstead of DMF Et₃N eq Heated 80° C for 4 hours then RT O/N Purified byHPLC Cis/Trans isomers separated after final step [M + H]⁺ 448 R_(t)1.97 171

Procedure D followed by G then E [M + H]⁺ 365 R_(t) 0.34 172

B Purified by column chromatography (pet. ether-EtOAc (1:1)) [M + H]⁺414.13 R_(t) 3.05 173

B Purified by column chromatography (pet. ether-EtOAc (1:1)) [M + H]⁺432.12 R_(t) 3.12 174

B Purified by column chromatography (pet. ether-EtOAc (1:1)) [M + H]⁺448.06 R_(t) 3.33 175

B Purified by column chromatography (pet. ether-EtOAc (1:1)) [M + H]⁺450.08 R_(t) 3.29 176

B Purified by column chromatography (pet. ether-EtOAc (1:1)) [M + H]⁺480.05 R_(t) 3.18 177

A Starting amine prepared according to Procedure L HOAt instead of HOBtDMSO as solvent instead of DMF Et₃N 2 eq Purified by HPLC and formationof HCl salt [M + H]⁺ 447 R_(t) 2.01 178

B [M + H]⁺ 343.05 R_(t) 3.38 (polar method) 179

A Butyl- piperidin-4- ylamine prepared by Procedure N HOAt instead ofHOBt Purified by trituration with MeOH [M + H]⁺ 406 R_(t) 1.85 180

B [M + H]⁺ 371.09 R_(t) 3.27 (polar method) 181

B [M + H]⁺ 306.06 R_(t) 1.53 182

B [M + H]⁺ 403.98 R_(t) 2.78 183

B [M + H]⁺ 345.05 R_(t) 3.03 184

B [M + H]⁺ 280.05 R_(t) 3.75 (basic method) 185

A HOAt instead of HOBt followed by EtOAc/HCl deprotection [M + H]⁺ 336R_(t) 1.67 186

A [M + H]⁺ 380.05 R_(t) 1.78 187

A [M + H]⁺ 396.02 R_(t) 1.86 188

A [M + H]⁺ 386.10 R_(t) 1.88 189

A [M + H]⁺ 342.10 R_(t) 1.95 190

M [M + H]⁺ = 344 R_(t) = 1.87 191

M [M + H]⁺ = 330 R_(t) = 1.80 192

M [M + H]⁺ = 372 R_(t) = 1.87 193

M [M + H]⁺ = 354 R_(t) = 1.77 194

M Purified by flash chromatography eluting with dichloromethane 120 ml,methanol 15, acetic acid 3 ml, water 2 ml (DMAW 120) [M + H]⁺ = 383/385R_(t) = 1.72 195

M Purified by flash chromatography eluting with DMAW 120 [M + H]⁺ =393/395 R_(t) = 1.86 196

M [M + H]⁺ = 398 R_(t) = 1.94 197

M [M + H]⁺ = 330 R_(t) = 1.80 198

M [M + H]⁺ = 358 R_(t) = 1.89 199

M [M + H]⁺ = 399 R_(t) = 1.88 200

M [M + H]⁺ = 420 R_(t) = 2.13 201

M [M + H]⁺ = 392/394 R_(t) = 1.84 202

B Purified using flash chromatography (CH₂Cl₂—MeOH—AcOH—H₂O (90:18:3:2))[M + H]⁺ 376.14 R_(t) 1.78 203

B Purified using flash chromatography (CH₂Cl₂—MeOH—AcOH—H₂O (90:18:3:2))[M + H]⁺ 400.17 R_(t) 2.08 204

B Purified using flash chromatography (CH₂Cl₂—MeOH—AcOH—H₂O (90:18:3:2))[M + H]⁺ 376.15 R_(t) 1.92 205

B Purified using column chromatography (CH₂Cl₂—MeOH—AcOH—H₂O(90:18:3:2)) [M + H]⁺ 382.12 R_(t) 1.77 206

B Purified using column chromatography (CH₂Cl₂—MeOH—AcOH—H₂O(90:18:3:2)) [M + H]⁺ 388.18 R_(t) 1.73 207

A Purified by flash chromatography eluting with DMAW 120 [M + H]⁺ =397/399 R_(t) = 1.83 208

A Coupling using (S)-3-amino-1- N-BOC-piperidine. Deprotection asprocedure M. Purified using column chromatography (CH₂Cl₂—MeOH—AcOH—H₂O(90:18:3:2)) [M + H]⁺ 382.02 R_(t) 1.82 209

A [M + H]⁺ 440.22 R_(t) 1.92 210

A [M + H]⁺ 411.20 R_(t) 2.97 211

A Purified by prep. LCMS after work-up [M + H]⁺ 362.11 R_(t) 1.91 212

A Purified by prep. LCMS after work-up [M + H]⁺ 396.08 R_(t) 2.06 213

A Purified by prep. LCMS after work-up [M + H]⁺ 396.06 R_(t) 2.04 214

B The mixture was reduced in vacuo, the residue taken up in EtOAc andwashed successively with saturated aqueous sodium biscarbonate, waterand brine. The organic portion was dried (MgSO₄) and reduced in vacuo togive the desired product [M + H]⁺ 485 R_(t) 2.59 215

B The mixture was reduced in vacuo, the residue taken up in EtOAc andwashed successively with saturated aqueous sodium biscarbonate, waterand brine. The organic portion was dried (MgSO₄) and reduced in vacuo togive the desired product [M + H]⁺ 429 R_(t) 2.25 216

A Purified by flash chromatography eluting with DMAW 120 [M + H]⁺ = 376R_(t) = 1.85 217

A Purified by flash chromatography eluting with DMAW 120 [M + H]⁺ = 376R_(t) = 1.87 218

A Purified by flash chromatography eluting with 5% then 10% MeOH/ DCM[M + H]⁺ = 376/378 R_(t) = 2.23 219

A Starting amine prepared according to Procedure L Purified by flashchromatography eluting with DMAW 90 [M + H]⁺ = 466/468 R_(t) = 1.98 220

A Purified by flash chromatography eluting with 5% then 10% MeOH/ DCM[M + H]⁺ = 376/378 R_(t) = 2.09 221

A Starting amine prepared according to Procedure L Purified by flashchromatography eluting with DMAW 90 [M + H]⁺ = 434 R_(t) = 1.82 222

A Purified by flash chromatography eluting with 5% then 10% MeOH/ DCM[M + H]⁺ = 356 R_(t) = 2.11 223

A Purified by flash chromatography eluting with 5% then 10% MeOH/ DCM[M + H]⁺ = 344 R_(t) = 2.09

Example 224 4-(4-Methyl-piperazin-1-yl)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

Bis(2-chloroethyl)methylamine hydrochloride (97 mg; 0.5 mmol) was addedto a stirred solution of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide (100 mg; 0.45 mmol), tetrabutylammonium iodide(20 mg; 0.045 mmol) and diisopropyethylamine (200 ul) 1.13 mmol) in DMF(5 ml), and the resulting mixture was heated at 200° C. (100 W) for 30minutes in a CEM Discover™ microwave synthesiser. The DMF was removedunder vacuum, then purified by flash column chromatography, eluting withdichloromethane/methanol/acetic acid/water (90:18:3:2). Productcontaining fractions were combined and evaporated, treated with HCl inethyl acetate and then re-evaporated with toluene (2×20 ml) to give anoff white solid (27 mg). (LC/MS: R_(t) 1.64, [M+H]⁺ 378).

Example 225 4-Morpholin-4-yl-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The compound was prepared in a manner analogous to Example 224, butusing bis(2-chloroethyl)ether in place of bis(2-chloroethyl)methylaminehydrochloride. (LC/MS: R_(t) 2.48 [M+H]⁺ 291).

Example 226 4-(2,4-Dichloro-phenyl)-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide

226A. Preparation of 4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylicacid

A solution of 4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylic acidethyl ester (205 mg; 0.72 mmol) and lithium hydroxide monohydrate (125mg; 2.9 mmol) in 1:1 THF/water (10 ml) was heated at 60° C. overnight.The THF was removed by evaporation, the aqueous phase acidified with 1Mhydrochloric acid then extracted with ethyl acetate (20 ml). The ethylacetate layer was dried (MgSO₄), filtered and evaporated to give 200 mgof 4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylic acid. (LC/MS: [M+H]⁺256.85).

226B. Preparation of 4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylicacid 4-(4-methyl-piperazin-1-yl)-benzylamide

A solution of 4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylic acid (70mg; 0.27 mmol), 4-(4-methyl-piperazin-1-yl)-benzylamine (62 mg; 0.3mmol), EDAC (63 mg; 0.33 mmol) and HOBt (45 mg; 0.33 mmol) in 5 ml ofDMF was stirred at room temperature for 48 hours. The reaction wasevaporated and the residue partitioned between ethyl acetate and brine.The ethyl acetate layer was separated, dried (MgSO₄), filtered,evaporated then dried further under vacuum to give 34 mg of4-(2,4-dichloro-phenyl)-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide. (LC/MS: R_(t) 2.42 [M+H]⁺ 444).

Example 227 4-(2,4-Dichloro-phenyl)-1H-pyrazole-3-carboxylic acid4-methylsulphamoylmethyl-benzylamide

The title compound was prepared in a manner analogous to Example 226,but using (4-aminomethyl-phenyl)-N-methyl-methanesulphonamide as thestarting material. 6 mg of product were isolated as a white solid.(LC/MS: R_(t) 3.56 [M+H]⁺ 440).

Example 228 4-Phenyl-1H-pyrazole-3-carboxylic acid amide

228A. 2-Benzylidene-but-3-yne nitrile

To a solution of benzaldehyde (2 g; 18.9 mmol) and malononitrile (1.37g; 20.7 mmol) in ethanol (40 ml) was added 5 drops of piperidine and themixture was heated at reflux overnight. The reaction was cooled,evaporated then purified by flash column chromatography eluting with 1:9ethyl acetate/hexane and the product containing fractions combined andevaporated to give 930 mg of 2-benzylidene-but-3-yne nitrile.

228B. 4-phenyl-5-trimethylsilanyl-1H-pyrazole-3-carbonitrile

n-Butyl lithium (2.7 M solution in heptane) (3.3 ml, 9 mmol) was addeddrop wise to a stirred solution of trimethylsilyl diazomethane (2 Msolution in diethyl ether) (4.5 ml, 9 mmol) in anhydrous THF (10 ml) at−78° C. under a nitrogen atmosphere, then stirred for a further 30minutes. To this was added drop wise a solution of2-benzylidene-but-3-yne nitrile (920 mg; 6 mmol) in anhydrous THF (5ml), the mixture stirred for 30 minutes at −78° C. then graduallyallowed to warm to room temperature overnight. The reaction mixture wasdiluted with ethyl acetate (30 ml) then washed with saturated ammoniumchloride solution followed by brine. The ethyl acetate layer wasseparated, dried (MgSO₄), filtered and evaporated. The crude product waspurified by flash column chromatography eluting with 1:8 then 1:4 ethylacetate/hexane and the product containing fractions combined andevaporated to give 1.0 g of4-phenyl-5-trimethylsilanyl-1H-pyrazole-3-carbonitrile.

228C. 4-phenyl-1H-pyrazole-3-carboxylic acid amide

4-Phenyl-5-trimethylsilanyl-1H-pyrazole-3-carbonitrile (500 mg; 2.1mmol) was dissolved in 1 ml of ethanol, treated with potassium hydroxide(600 mg) in water (3 ml) then heated at 150° C. (100 W) for 30 minutesthen 170° C. (100 W) for 20 minutes in a CEM Discover™ microwavesynthesiser. The reaction mixture was acidified to pH1 with concentratedhydrochloric acid, diluted with water (40 ml) then extracted with ethylacetate (2×40 ml). The combined ethyl acetate layers were separated,dried (MgSO₄), filtered and evaporated to give a 3:1 mixture of4-phenyl-1H-pyrazole-3-carboxylic acid and4-phenyl-1H-pyrazole-3-carboxylic acid amide. A 50 mg batch of the crudematerial was purified by flash column chromatography eluting with 5%methanol/dichloromethane, and the product containing fractions combinedand evaporated to give 15 mg of 4-phenyl-1H-pyrazole-3-carboxylic acidamide as a white solid. (LC/MS: R_(t) 2.15 [M+H]⁺ 188).

Example 229 4-phenyl-1H-pyrazole-3-carboxylic acid phenylamide

A solution of 4-phenyl-1H-pyrazole-3-carboxylic acid (75 mg; 0.4 mmol)(prepared according to Example 228C), aniline (45 μA; 0.48 mmol), EDAC(92 mg; 0.48 mmol) and HOBt (65 mg; 0.48 mmol) in 5 ml of DMF wasstirred at room temperature overnight. The reaction was evaporated thenpurified by flash column chromatography eluting with 1:3 then 1:2 ethylacetate/hexane. Product containing fractions were combined andevaporated to give 30 mg of 4-phenyl-1H-pyrazole-3-carboxylic acidphenylamide as a white solid. (LC/MS: R_(t) 3.12 [M+H]⁺ 264).

Example 230 4-Phenyl-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide

The compound was prepared in a manner analogous to Example 229, butusing 4-(4-methyl-piperazin-1-yl)-benzylamine as the starting material.6 mg of product were isolated as a white solid. (LC/MS: R_(t) 2.05[M+H]⁺ 376).

Example 231 4-Phenyl-1H-pyrazole-3-carboxylic acid(6-methoxy-pyridin-3-yl)amide

The compound was prepared in a manner analogous to Example 230, butusing 3-amino-6-methoxypyridine as the amine fragment. 100 mg of productwere isolated as a pale brown solid. (LC/MS: R_(t) 3.17 [M+H]⁺ 295).

Example 232 4-(3-Benzyloxy-phenyl)-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide

The compound was prepared in a manner analogous to Example 226. Theproduct was isolated as a white solid. (LC/MS: R_(t) 2.65 [M+H]⁺482).

Example 233 4-(3-Hydroxy-phenyl)-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide

A solution of 4-(3-benzyloxy-phenyl)-1H-pyrazole-3-carboxylic acid4-(4-methyl-piperazin-1-yl)-benzylamide (25 mg; 0.05 mmol) in methanol(5 ml), was treated with 10% palladium on carbon (10 mg) thenhydrogenated at room temperature and pressure overnight. The catalystwas removed by filtration through Celite and the filtrate evaporated.Purification by preparative LC/MS gave 8 mg of the required product as acream solid. (LC/MS: R_(t) 1.67 [M+H]⁺ 392).

Example 234 4-(5-Methyl-3H-imidazol-4-yl)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

The compound was prepared in a manner analogous to Example 226, butusing 4-methyl-5-formylimidazole as the starting material in thecondensation step. The product (6 mg) was isolated as a white solid.(LC/MS: R_(t) 2.00 [M+H]⁺ 286).

Example 235 4-(2,5-Dimethyl-pyrrol-1-yl)-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

A mixture of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide (100 mg) and Montmorillonite KSF clay (100 mg)in acetonylacetone (1 ml) was heated at 120° C. (50 W) for 15 minutes ina CEM discover microwave synthesiser. The reaction mixture was dilutedwith 5% methanol/dichloromethane, filtered and evaporated. The crudeproduct was purified by flash column chromatography eluting with 1:2ethyl acetate/hexane, and the product containing fractions were combinedand evaporated to give 65 mg of the target molecule as a pale brownsolid. (LC/MS: R_(t) 3.75 [M+H]⁺ 299).

Example 236 4-(3-Hydroxymethyl-phenyl)-1H-pyrazole-3-carboxylic acidphenylamide

236A. 4-iodo-1H-pyrazole-3-carboxylic acid phenylamide

An aqueous solution of sodium nitrite (760 mg) in 2 ml of water wasadded drop wise to a stirred suspension of4-amino-1H-pyrazole-3-carboxylic acid phenylamide (2 g; 10 mmol) inconcentrated hydrochloric acid (20 ml) at 0° C., then stirred at 0° C.for a further 60 minutes. The reaction mixture was diluted with acetone(10 ml) then treated with potassium iodide (1.8 g) and copper (I) iodide(2.1 g) and stirred at room temperature for 90 minutes. The reactionmixture was diluted with brine and ethyl acetate then washed withsaturated sodium thiosulphate solution. The ethyl acetate layer wasseparated, dried (MgSO₄), filtered and evaporated to give 680 mg of4-iodo-1H-pyrazole-3-carboxylic acid phenylamide.

236B. 4-iodo-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylic acidphenylamide

A solution of 4-iodo-1H-pyrazole-3-carboxylic acid phenylamide (670 mg;2.14 mmol) in acetonitrile (10 ml) was treated with potassium carbonate(360 mg; 2.57 mmol)) followed by 4-methoxybenzyl chloride (320 μl; 2.35mmol). The mixture was stirred at room temperature overnight thenevaporated under reduced pressure. The residue was partitioned betweenethyl acetate and brine; the ethyl acetate layer was separated, dried(MgSO₄), filtered and evaporated. The crude material was purified byflash column chromatography eluting with 1:3 ethyl acetate/hexane andthe product containing fractions combined and evaporated to give 660 mgof 4-iodo-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylic acidphenylamide.

236C.4-(3-hydroxymethyl-phenyl)-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylicacid phenylamide

A mixture of 4-iodo-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylic acidphenylamide (50 mg; 0.11 mmol), bis(tri-tert-butylphosphine)palladium(12 mg), potassium carbonate (100 mg; 0.66 mmol) and3-(hydroxmethyl)benzene boronic acid (21 mg; 0.14 mmol) inethanol/toluene/water (4 ml:1 ml:1 ml) was heated at 120° C. (50 W) for15 minutes in a CEM Discover microwave synthesiser. The reaction wasevaporated and the residue partitioned between ethyl acetate and brine.The ethyl acetate layer was separated, dried (MgSO₄), filtered andevaporated and the crude material purified by flash columnchromatography eluting with 1:2 then 2:1 ethyl acetate/hexane. Productcontaining fractions were combined and evaporated to give 60 mg of4-(3-hydroxymethyl-phenyl)-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylicacid phenylamide.

236D. 4-(3-Hydroxymethyl-phenyl)-1H-pyrazole-3-carboxylic acidphenylamide

A mixture of4-(3-hydroxymethyl-phenyl)-1-(4-methoxy-benzyl)-1H-pyrazole-3-carboxylicacid phenylamide (20 mg) and anisole (20 μl) in trifluoroacetic acid (1ml) was heated at 120° C. (50 W) for 15 minutes in a CEM Discovermicrowave synthesiser. The reaction was evaporated then purified byflash column chromatography eluting with 2:1 ethyl acetate/hexane.Product containing fractions were combined and evaporated to give 5 mgof product. (LC/MS: R_(t) 2.55 [M+H]⁺ 294).

Example 237 Preparation of4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide hydrochloride 237A.4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid

2,6-dichlorobenzoyl chloride (8.2 g; 39.05 mmol) was added cautiously toa solution of 4-amino-1H-pyrazole-3-carboxylic acid methyl ester(prepared in a manner analogous to 165B) (5 g; 35.5 mmol) andtriethylamine (5.95 ml; 42.6 mmol) in dioxan (50 ml) then stirred atroom temperature for 5 hours. The reaction mixture was filtered and thefiltrate treated with methanol (50 ml) and 2M sodium hydroxide solution(100 ml), heated at 50° C. for 4 hours, and then evaporated. 100 ml ofwater was added to the residue then acidified with concentratedhydrochloric acid. The solid was collected by filtration, washed withwater (100 ml) and sucked dry to give 10.05 g of4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid as a paleviolet solid.

237B.4-{[4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-piperidine-1-carboxylicacid tert-butyl ester

A mixture of 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid(6.5 g, 21.6 mmol), 4-amino-1-BOC-piperidine (4.76 g, 23.8 mmol), EDC(5.0 g, 25.9 mmol) and HOBt (3.5 g, 25.9 mmol) in DMF (75 ml) wasstirred at room temperature for 20 hours. The reaction mixture wasreduced in vacuo and the residue partitioned between ethyl acetate (100ml) and saturated aqueous sodium bicarbonate solution (100 ml). Theorganic layer was washed with brine, dried (MgSO₄) and reduced in vacuo.The residue was taken up in 5% MeOH-DCM (˜30 ml). The insoluble materialwas collected by filtration and, washed with DCM and dried in vacuo togive4-{[4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-piperidine-1-carboxylicacid tert-butyl ester (5.38 g) as a white solid. The filtrate wasreduced in vacuo and the residue purified by column chromatography usinggradient elution 1:2 EtOAc/hexane to EtOAc to give further4-{[4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-piperidine-1-carboxylicacid tert-butyl ester (2.54 g) as a white solid.

237C. 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide

A solution of4-{[4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carbonyl]-amino}-piperidine-1-carboxylicacid tert-butyl ester (7.9 g) in MeOH (50 mL) and EtOAc (50 ml) wastreated with sat. HCl-EtOAc (40 mL) then stirred at r.t. overnight. Theproduct did not crystallise due to the presence of methanol, andtherefore the reaction mixture was evaporated and the residue trituratedwith EtOAc. The resulting off white solid was collected by filtration,washed with EtOAc and sucked dry on the sinter to give 6.3 g of4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide as the hydrochloride salt. (LC/MS: R_(t) 5.89,[M+H]⁺ 382/384).

Example 238 4-Methanesulfonylamino-1H-pyrazole-3-carboxylic acid(4-fluoro-phenyl)-amide

A solution of 4-amino-1H-pyrazole-3-carboxylic acid(4-fluorophenyl)-amide (50 mg) (Example 2B) and methanesulphonicanhydride (45 mg) in pyridine (1 ml) was stirred at room temperatureovernight then evaporated and purified by flash column chromatographyeluting with 2:1 EtOAc/hexane. Evaporation of product containingfractions gave 20 mg of the title compound. (LC/MS: R_(t) 2.87; [M+H+]299).

Examples 239 to 245

The compounds of Examples 239 to 245 were prepared using the methodsdescribed above or methods closely analogous thereto.

Example 239 4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[1-(2-fluoro-ethyl)-piperidin-4-yl]-amide

Example 240 4-(2,6-Dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid(6-chloro-pyridin-3-yl)-amide

Example 241 4-(2,6-Dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid(6-amino-pyridin-3-yl)-amide

Example 242 4-(2,6-Dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acid(6-methoxy-pyridin-3-yl)-amide

Example 2434-[3-Chloro-5-(4-methyl-piperazin-1-yl)-benzoylamino]-1H-pyrazole-3-carboxylicacid cyclohexylamide

Example 244 4-(2,6-Difluoro-benzoylamino)-1H-pyrazole-3-carboxylic acid[1-(2,2-difluoro-ethyl)-piperidin-4-yl]-amide

Example 2454-[3-(4-Methyl-piperazin-1-yl)-benzoylamino]-1H-pyrazole-3-carboxylicacid cyclohexylamide

Biological Activity Example 246 Measurement of CDK2 Kinase InhibitoryActivity (IC₅₀)

Compounds of the invention were tested for kinase inhibitory activityusing either the following protocol or the activated CDK2/cyclin Akinase protocol described in Example 241.

1.7 μl of active CDK2/CyclinA (Upstate Biotechnology, 10 U/□μl) isdiluted in assay buffer (250□μl of 10× strength assay buffer (200 mMMOPS pH 7.2, 250 mM β-glycerophosphate, 50 mM EDTA, 150 mM MgCl₂), 11.27μl 10 mM ATP, 2.5 μl 1M DTT, 25 μl 100 mM sodium orthovanadate, 708.53μl H₂O), and 10 μl mixed with 10 μl of histone substrate mix (60 μlbovine histone H1 (Upstate Biotechnology, 5 mg/ml), 940 μl H₂O, 35 μCiγ³³P-ATP) and added to 96 well plates along with 5 μl of variousdilutions of the test compound in DMSO (up to 2.5%). The reaction isallowed to proceed for 5 hours before being stopped with an excess ofortho-phosphoric acid (30 μl at 2%).

γ³³P-ATP which remains unincorporated into the histone H1 is separatedfrom phosphorylated histone H1 on a Millipore MAPH filter plate. Thewells of the MAPH plate are wetted with 0.5% orthophosphoric acid, andthen the results of the reaction are filtered with a Millipore vacuumfiltration unit through the wells. Following filtration, the residue iswashed twice with 200 μl of 0.5% orthophosphoric acid. Once the filtershave dried, 25 μl of Microscint 20 scintillant is added, and thencounted on a Packard Topcount for 30 seconds.

The % inhibition of the CDK2 activity is calculated and plotted in orderto determine the concentration of test compound required to inhibit 50%of the CDK2 activity (IC₅₀).

By means of the protocol set out above, it was found that the compoundsof Examples 2C to 87, 89-92, 94, 96-101, 104-105, 165, 166, 224, 225,227, 229, 231, 233, 234 and 236 each have IC₅₀ values less than 20 μM orprovide at least 50% inhibition of the CDK2 activity at a concentrationof 10 μM. The compounds of Examples 88, 93, 226, 228, 230 and 235 eachhave IC₅₀ values less than 750 μM.

Example 247 CDK Selectivity Assays

Compounds of the invention are tested for kinase inhibitory activityagainst a number of different kinases using the general protocoldescribed in Example 239, but modified as set out below.

Kinases are diluted to a 10× working stock in 20 mM MOPS pH 7.0, 1 mMEDTA, 0.1% γ-mercaptoethanol, 0.01% Brij-35, 5% glycerol, 1 mg/ml BSA.One unit equals the incorporation of 1 nmol of phosphate per minute into0.1 mg/ml histone H1, or CDK7 substrate peptide at 30□° C. with a finalATP concentration of 100 μM.

The substrate for all the CDK assays (except CDK7) is histone H1,diluted to 10× working stock in 20 mM MOPS pH 7.4 prior to use. Thesubstrate for CDK7 is a specific peptide obtained from Upstate dilutedto 10× working stock in deionised water.

Assay Procedure for CDK1/cyclinB, CDK2/cyclinA, CDK2/cyclinE,CDK3/cyclinE, CDK5/p35, CDK6/cyclinD3:

In a final reaction volume of 25 μl, the enzyme (5-10 mU) is incubatedwith 8 mM MOPS pH 7.0, 0.2 mM EDTA, 0.1 mg/ml histone H1, 10 mMMgAcetate and [γ-³³P-ATP] (specific activity approx 500 cpm/pmol,concentration as required). The reaction is initiated by the addition ofMg²⁺ [γ-³³P-ATP]. After incubation for 40 minutes at room temperaturethe reaction is stopped by the addition of 5 μl of a 3% phosphoric acidsolution. 10 ml of the reaction is spotted onto a P30 filter mat andwashed 3 times for 5 minutes in 75 mM phosphoric acid and once inmethanol prior to drying and counting.

In the CDK3/cyclinE assay, the compound of Example 150 had an IC₅₀ ofless than 20 μM.

In the CDK5/p35 assay, the compounds of Examples 41 and 150 had an IC₅₀of less than 20 μM.

In the CDK6/cyclinD3 assay, the compound of Example 150 had an IC₅₀ ofless than 20 μM.

Assay Procedure for CDK7/cyclinH/MAT1

In a final reaction volume of 25 μl, the enzyme (5-10 mU) is incubatedwith 8 mM MOPS pH 7.0, 0.2 mM EDTA, 500 μM peptide, 10 mM MgAcetate and[γ-³³P-ATP] (specific activity approx 500 cpm/pmol, concentration asrequired). The reaction is initiated by the addition of Mg²⁺[γ-³³P-ATP].After incubation for 40 minutes at room temperature the reaction isstopped by the addition of 5 μl of a 3% phosphoric acid solution. 10 mlof the reaction is spotted onto a P30 filtermat and washed 3 times for 5minutes in 75 mM phosphoric acid and once in methanol prior to dryingand counting.

Example 248 A. Measurement of Activated CDK2/CyclinA Kinase InhibitoryActivity Assay (IC₅₀)

Compounds of the invention were tested for kinase inhibitory activityusing the following protocol.

Activated CDK2/CyclinA (Brown et al, Nat. Cell Biol., 1, pp 438-443,1999; Lowe, E. D., et al Biochemistry, 41, pp 15625-15634, 2002) isdiluted to 125 μM in 2.5× strength assay buffer (50 mM MOPS pH 7.2, 62.5mM β-glycerophosphate, 12.5 mM EDTA, 37.5 mM MgCl₂, 112.5 mM ATP, 2.5 mMDTT, 2.5 mM sodium orthovanadate, 0.25 mg/ml bovine serum albumin), and10 μl mixed with 10 μl of histone substrate mix (60 μl bovine histone H1(Upstate Biotechnology, 5 mg/ml), 940 μl H₂O, 35 μCi γ³³P-ATP) and addedto 96 well plates along with 5 μl of various dilutions of the testcompound in DMSO (up to 2.5%). The reaction is allowed to proceed for 2to 4 hours before being stopped with an excess of ortho-phosphoric acid(5 μl at 2%).

γ³³P-ATP which remains unincorporated into the histone H1 is separatedfrom phosphorylated histone H1 on a Millipore MAPH filter plate. Thewells of the MAPH plate are wetted with 0.5% orthophosphoric acid, andthen the results of the reaction are filtered with a Millipore vacuumfiltration unit through the wells. Following filtration, the residue iswashed twice with 200 μl of 0.5% orthophosphoric acid. Once the filtershave dried, 20 μl of Microscint 20 scintillant is added, and thencounted on a Packard Topcount for 30 seconds.

The % inhibition of the CDK2 activity is calculated and plotted in orderto determine the concentration of test compound required to inhibit 50%of the CDK2 activity (IC₅₀).

By means of the foregoing protocol, it was found that the compounds ofExamples 95, 96, 99-104, 106-121, 123-125, 130-137, 139, 142-145,147-150, 152-156, 158-160, 162-164, 167-173, 177-179, 181-182, 184-190,194, 196-204, 208-213 and 215 have IC₅₀ values less than 20 μM. Thecompounds of Examples 122, 126-129, 140, 141, 146, 157 and 161 each haveIC₅₀ values less than 750 μM and most have IC₅₀ values of less than 100μM.

B. CDK1/CyclinB Assay

CDK1/CyclinB assay is identical to the CDK2/CyclinA above except thatCDK1/CyclinB (Upstate Discovery) is used and the enzyme is diluted to6.25 nM.

In the CDK1 assay carried out as described above or by means of theprotocol set out in Example 240, the compounds of Examples 2C, 41, 48,53, 64, 65, 66, 73, 76, 77, 91, 95, 102, 106, 117, 123, 125, 133, 137,142, 150, 152, 154, 167, 186, 187, 189, 190, 193, 194, 196, 199,202-204, 207, 208-213, 215 AND 218-223 were found to have IC₅₀ valuesless than 20 and the compounds of Examples 188 and 206, were found tohave IC₅₀ values less than 100 μM.

Example 249 Assay Procedure for CDK4

Assays for CDK4 inhibitory activity were carried out by Proqinase GmbH,Freiburg, Germany using their proprietary 33 PanQinase® Activity Assay.The assays were performed in 96 well FlashPlates™ (PerkinElmer). In eachcase, the reaction cocktail (50 μl final volume) is composed of; 20 μlassay buffer (final composition 60 mM HEPES-NaOH, pH 7.5, 3 mM MgCl₂, 3μM Na-orthovanadate, 1.2 mM DTT, 50□μg/ml PEG₂₀₀₀, 5□μl ATP solution(final concentration 1 μM [γ-33P]-ATP (approx 5×10⁵ cpm per well)), 5□μltest compound (in 10% DMSO), 10□μl substrate/10□μl enzyme solution(premixed). The final amounts of enzyme and substrate were as below.

Kinase ng/ Substrate ng/ Kinase 50□ μl Substrate 50□ μl CDK4/CycD1 50Poly (Ala, Glu, Lys, 500 Tyr) 6:2:5:1

The reaction cocktail was incubated at 30° C. for 80 minutes. Thereaction was stopped with 50 μl of 2% H₃PO₄, plates were aspirated andwashed twice with 200 μl 0.9% NaCl. Incorporation of ³³P was determinedwith a microplate scintillation counter. Background values weresubtracted from the data before calculating the residual activities foreach well. IC₅₀s were calculated using Prism 3.03.

The compound of Example 150 has an IC50 of less than 5 μM in this assay.

Example 250 Anti-Proliferative Activity

The anti-proliferative activities of compounds of the invention aredetermined by measuring the ability of the compounds to inhibition ofcell growth in a number of cell lines. Inhibition of cell growth ismeasured using the Alamar Blue assay (Nociari, M. M, Shalev, A., Benias,P., Russo, C. Journal of Immunological Methods 1998, 213, 157-167). Themethod is based on the ability of viable cells to reduce resazurin toits fluorescent product resorufin. For each proliferation assay cellsare plated onto 96 well plates and allowed to recover for 16 hours priorto the addition of inhibitor compounds for a further 72 hours. At theend of the incubation period 10% (v/v) Alamar Blue is added andincubated for a further 6 hours prior to determination of fluorescentproduct at 535 nM ex/590 nM em. In the case of the non-proliferatingcell assay cells are maintained at confluence for 96 hour prior to theaddition of inhibitor compounds for a further 72 hours. The number ofviable cells is determined by Alamar Blue assay as before. All celllines are obtained from ECACC (European Collection of cell Cultures).

In assays against the human colon carcinoma cell line HCT 116 (ECACC No.91091005), the compounds of Examples 10, 25-27, 41, 44, 46, 48, 50, 52,53, 60, 62, 64-67, 69, 73-77, 79, 80, 83A, 86, 90-93, 95-98, 100-104,106, 107, 109-121, 123-125, 131-134, 136-143, 147-155, 158, 159,162-164, 166, 167, 178, 179, 185-190, 192-205, 207-215 and 218-223 haveIC₅₀ values of less than 20 μM and the compounds of Examples 2C, 3, 29,38, 39, 49, 51, 85, 89, 99, 108, 135, 160, 182, 183, 206 and 216 haveIC₅₀ values of less than 100 μM.

Example 251 Measurement of Inhibitory Activity Against Glycogen SynthaseKinase-3 (GSK-3)

The activities of the compounds of the invention as inhibitors of GSK-3were determined using either Protocol A or Protocol B below.

Protocol A

GSK3-β (Upstate Discovery) is diluted to 7.5 nM in 25 mM MOPS, pH 7.00,25 mg/ml BSA, 0.0025% Brij-35®, 1.25% glycerol, 0.5 mM EDTA, 25 mMMgCl₂, 0.025% β-mercaptoethanol, 37.5 mM ATP and 10 μl mixed with 10 μlof substrate mix. The substrate mix is 12.5 μM phospho-glycogen synthasepeptide-2 (Upstate Discovery) in 1 ml of water with 35 μCi γ³³P-ATP.Enzyme and substrate are added to 96 well plates along with 5 μl ofvarious dilutions of the test compound in DMSO (up to 2.5%). Thereaction is allowed to proceed for 3 hours before being stopped with anexcess of ortho-phosphoric acid (5 μl at 2%). The filtration procedureis as for Activated CDK2/CyclinA assay above.

Protocol B

GSK3β (human) is diluted to a 10× working stock in 50 mM Tris pH 7.5,0.1 mM EGTA, 0.1 mM sodium vanadate, 0.1% β-mercaptoethanol, 1 mg/mlBSA. One unit equals the incorporation of 1 nmol of phosphate per minutephospho-glycogen synthase peptide 2 per minute.

In a final reaction volume of 25 μl, GSK3β (5-10 mU) is incubated with 8mM MOPS 7.0, 0.2 mM EDTA, 20 μM YRRAAVPPSPSLSRHSSPHQS(p)EDEEE (phosphoGS2 peptide), 10 mM MgAcetate and [ε-³³P-ATP] (specific activity approx500 cpm/pmol, concentration as required). The reaction is initiated bythe addition of Mg²⁺[γ-³³P-ATP]. After incubation for 40 minutes at roomtemperature the reaction is stopped by the addition of 5 μl of a 3%phosphoric acid solution. 10 μl of the reaction is spotted onto a P30filter mat and washed 3 times for 5 minutes in 50 mM phosphoric acid andonce in methanol prior to drying and counting.

From the results of the GSK3-B assays carried out using either of thetwo protocols set out above, it was found that the compounds of Examples2C, 26, 48, 53, 65, 76, 77, 84, 86, 95, 102, 106, 119, 122, 123, 126,127, 128, 129, 131, 134, 135, 138, 140, 141, 142, 143, 144, 145, 146,147, 149, 150 and 151 each have IC₅₀ values of less than 10 μM.

Pharmaceutical Formulations Example 252 (i) Tablet Formulation

A tablet composition containing a compound of the formula (I) isprepared by mixing 50 mg of the compound with 197 mg of lactose (BP) asdiluent, and 3 mg magnesium stearate as a lubricant and compressing toform a tablet in known manner.

(ii) Capsule Formulation

A capsule formulation is prepared by mixing 100 mg of a compound of theformula (I) with 100 mg lactose and filling the resulting mixture intostandard opaque hard gelatin capsules.

(iii) Injectable Formulation I

A parenteral composition for administration by injection can be preparedby dissolving a compound of the formula (I) (e.g. in a salt form) inwater containing 10% propylene glycol to give a concentration of activecompound of 1.5% by weight. The solution is then sterilised byfiltration, filled into an ampoule and sealed.

(iv) Injectable Formulation II

A parenteral composition for injection is prepared by dissolving inwater a compound of the formula (I) (e.g. in salt form) (2 mg/ml) andmannitol (50 mg/ml), sterile filtering the solution and filling intosealable 1 ml vials or ampoules.

(iv) Subcutaneous Injection Formulation

A composition for sub-cutaneous administration is prepared by mixing acompound of the formula (I) with pharmaceutical grade corn oil to give aconcentration of 5 mg/ml. The composition is sterilised and filled intoa suitable container.

Example 253 Determination of Antifungal Activity

The antifungal activity of the compounds of the formula (I) isdetermined using the following protocol.

The compounds are tested against a panel of fungi including Candidaparpsilosis, Candida tropicalis, Candida albicans-ATCC 36082 andCryptococcus neoformans. The test organisms are maintained on SabourandDextrose Agar slants at 4° C. Singlet suspensions of each organism areprepared by growing the yeast overnight at 27° C. on a rotating drum inyeast-nitrogen base broth (YNB) with amino acids (Difco, Detroit,Mich.), pH 7.0 with 0.05 morpholine propanesulphonic acid (MOPS). Thesuspension is then centrifuged and washed twice with 0.85% NaCl beforesonicating the washed cell suspension for 4 seconds (Branson Sonifier,model 350, Danbury, Conn.). The singlet blastospores are counted in ahaemocytometer and adjusted to the desired concentration in 0.85% NaCl.

The activity of the test compounds is determined using a modification ofa broth microdilution technique. Test compounds are diluted in DMSO to a1.0 mg/ml ratio then diluted to 64 μg/ml in YNB broth, pH 7.0 with MOPS(Fluconazole is used as the control) to provide a working solution ofeach compound. Using a 96-well plate, wells 1 and 3 through 12 areprepared with YNB broth, ten fold dilutions of the compound solution aremade in wells 2 to 11 (concentration ranges are 64 to 0.125 μg/ml). Well1 serves as a sterility control and blank for the spectrophotometricassays. Well 12 serves as a growth control. The microtitre plates areinoculated with 10 μl in each of well 2 to 11 (final inoculum size is10⁴ organisms/ml). Inoculated plates are incubated for 48 hours at 35°C. The MIC values are determined spectrophotometrically by measuring theabsorbance at 420 nm (Automatic Microplate Reader, DuPont Instruments,Wilmington, Del.) after agitation of the plates for 2 minutes with avortex-mixer (Vorte-Genie 2 Mixer, Scientific Industries, Inc., Bolemia,N.Y.). The MIC endpoint is defined as the lowest drug concentrationexhibiting approximately 50% (or more) reduction of the growth comparedwith the control well. With the turbidity assay this is defined as thelowest drug concentration at which turbidity in the well is <50% of thecontrol (IC50). Minimal Cytolytic Concentrations (MCC) are determined bysub-culturing all wells from the 96-well plate onto a Sabourand DextroseAgar (SDA) plate, incubating for 1 to 2 days at 35° C. and then checkingviability.

Example 254 Protocol for the Biological Evaluation of Control of in vivoWhole Plant Fungal Infection

Compounds of the formula (I) are dissolved in acetone, with subsequentserial dilutions in acetone to obtain a range of desired concentrations.Final treatment volumes are obtained by adding 9 volumes of 0.05%aqueous Tween-20 ™ or 0.01% Triton X-100™, depending upon the pathogen.

The compositions are then used to test the activity of the compounds ofthe invention against tomato blight (Phytophthora infestans) using thefollowing protocol. Tomatoes (cultivar Rutgers) are grown from seed in asoil-less peat-based potting mixture until the seedlings are 10-20 cmtall. The plants are then sprayed to run-off with the test compound at arate of 100 ppm. After 24 hours the test plants are inoculated byspraying with an aqueous sporangia suspension of Phytophthora infestans,and kept in a dew chamber overnight. The plants are then transferred tothe greenhouse until disease develops on the untreated control plants.

Similar protocols are also used to test the activity of the compounds ofthe invention in combatting Brown Rust of Wheat (Puccinia), PowderyMildew of Wheat (Ervsiphe vraminis), Wheat (cultivar Monon), Leaf Blotchof Wheat (Septoria tritici), and Glume Blotch of Wheat (Leptosphaerianodorum).

EQUIVALENTS

-   1. The foregoing examples are presented for the purpose of    illustrating the invention and should not be construed as imposing    any limitation on the scope of the invention. It will readily be    apparent that numerous modifications and alterations may be made to    the specific embodiments of the invention described above and    illustrated in the examples without departing from the principles    underlying the invention. All such modifications and alterations are    intended to be embraced by this application.

1. A combination comprising: (i) a compound of the formula (II):

or salts or tautomers or N-oxides or solvates thereof; wherein Y is abond or an alkylene chain of 1, 2 or 3 carbon atoms in length; R¹ is acarbocyclic or heterocyclic group having from 3 to 12 ring members,wherein the carbocyclic or heterocyclic groups are unsubstituted orsubstituted by one or more substituent groups R¹⁰; or a C₁₋₈ hydrocarbylgroup optionally substituted by one or more substituents selected fromfluorine, hydroxy, C₁₋₄ hydrocarbyloxy, amino, mono- or di-C₁₋₄hydrocarbylamino, and carbocyclic or heterocyclic groups having from 3to 12 ring members wherein the carbocyclic or heterocyclic groups areunsubstituted or substituted by one or more substituent groups R¹⁰; R²is hydrogen or methyl; R³ is selected from non-aromatic carbocyclic andheterocyclic groups having from 3 to 12 ring members, wherein thecarbocyclic or heterocyclic groups are unsubstituted or substituted byone or more substituent groups R¹⁰; and R¹⁰ is selected from halogen,hydroxy, trifluoromethyl, cyano, nitro, carboxy, amino, mono- or di-C₁₋₄hydrocarbylamino, carbocyclic and heterocyclic groups having from 3 to12 ring members; a group R^(a)-R^(b) wherein R^(a) is a bond, O, CO,X¹C(X²), C(X²)X¹, X¹C(X²)X¹, S, SO, SO₂, NR^(c), SO₂NR^(c) or NR^(c)SO₂;and R^(b) is selected from hydrogen, carbocyclic and heterocyclic groupshaving from 3 to 12 ring members, and a C₁₋₈ hydrocarbyl groupoptionally substituted by one or more substituents selected fromhydroxy, oxo, halogen, cyano, nitro, carboxy, amino, mono- or di-C₁₋₄hydrocarbylamino, carbocyclic and heterocyclic groups having from 3 to12 ring members; R^(c) is selected from hydrogen and C₁₋₄ hydrocarbyl;and X¹ is O, S or NR^(c) and X² is ═O, ═S or ═NR^(c); and provided thatwhere the substituent group R¹⁰ comprises or includes a carbocyclic orheterocyclic group, the said carbocyclic or heterocyclic group may beunsubstituted or may itself be substituted with one or more furthersubstituent groups R¹⁰ and wherein (a) such further substituent groupsR¹⁰ include carbocyclic or heterocyclic groups, which are not themselvesfurther substituted; or (b) the said further substituents do not includecarbocyclic or heterocyclic groups but are otherwise selected from thegroups listed above in the definition of R¹⁰; and (ii) one or more othertherapeutic agents.
 2. A combination according to claim 1 wherein R² ishydrogen.
 3. A combination according to claim 1 wherein R¹ is anunsubstituted or substituted monocyclic carbocyclic or heterocyclicgroup wherein the carbocyclic and heterocyclic groups are substituted byone or more substituent groups R¹⁰ or R^(10a); wherein R¹⁰ is as definedin claim 1 and R^(10a) is selected from halogen, hydroxy,trifluoromethyl, cyano, nitro, carboxy, a group R^(a)-R^(b) whereinR^(a) is a bond, O, CO, X³C(X⁴), C(X⁴)X³, X³C(X⁴)X³, S, SO, or SO₂, andR^(b) is selected from hydrogen and a C₁₋₈ hydrocarbyl group optionallysubstituted by one or more substituents selected from hydroxy, oxo,halogen, cyano, nitro, carboxy and monocyclic non-aromatic carbocyclicor heterocyclic groups having from 3 to 6 ring members; wherein one ormore carbon atoms of the C₁₋₈ hydrocarbyl group may optionally bereplaced by O, S, SO, SO₂, X³C(X⁴), C(X⁴)X³ or X³C(X⁴)X³; X³ is O or S;and X⁴ is ═O or ═S.
 4. A combination according to claim 1 wherein thecompound of formula (II) is of the formula (IV):

or salts or tautomers or N-oxides or solvates thereof; wherein R¹ and R²are as defined in any one of the preceding claims; an optional secondbond may be present between carbon atoms numbered 1 and 2; one of U andT is selected from CH₂, CHR¹³, CR¹¹R¹³, NR¹⁴, N(O)R¹⁵, O and S(O)_(t);and the other of U and T is selected from, NR¹⁴, O, CH₂, CHR¹¹, C(R¹¹)₂,and C═O; r is 0, 1, 2, 3 or 4; t is 0, 1 or 2; R¹¹ is selected fromhydrogen, halogen, C₁₋₃ alkyl and C₁₋₃ alkoxy; R¹³ is selected fromhydrogen, NHR¹⁴, NOH, NOR¹⁴ and R^(a)-R^(b); R¹⁴ is selected fromhydrogen and R^(d)-R^(b); R^(d) is selected from a bond, CO, C(X²)X¹,SO₂ and SO₂NR^(c); R^(a), R^(b) and R^(c) are as defined in any one ofthe preceding claims; and R¹⁵ is selected from C₁₋₄ saturatedhydrocarbyl optionally substituted by hydroxy, C₁₋₂ alkoxy, halogen or amonocyclic 5- or 6-membered carbocyclic or heterocyclic group, providedthat U and T cannot be O simultaneously.
 5. A combination according toclaim 4 wherein the compound of formula (II) is of the formula (Va):

or salts or tautomers or N-oxides or solvates thereof; wherein R^(14a)is selected from hydrogen, C₁₋₄ alkyl optionally substituted by fluoro,cyclopropylmethyl, phenyl-C₁₋₂ alkyl, C₁₋₄ alkoxycarbonyl, phenyl-C₁₋₂alkoxycarbonyl, C₁₋₂-alkoxy-C₁₋₂ alkyl, and C₁₋₄ alkylsulphonyl, whereinthe phenyl moieties when present are optionally substituted by one tothree substituents selected from fluorine, chlorine, C₁₋₄ alkoxyoptionally substituted by fluoro or C₁₋₂-alkoxy, and C₁₋₄ alkyloptionally substituted by fluoro or C₁₋₂-alkoxy; w is 0, 1, 2 or 3; R²is hydrogen or methyl; R¹¹ and r are as defined in any one of claims 17to 19; and R¹⁹ is selected from fluorine; chlorine; C₁₋₄ alkoxyoptionally substituted by fluoro or C₁₋₂-alkoxy; and C₁₋₄ alkyloptionally substituted by fluoro or C₁₋₂-alkoxy.
 6. A combinationaccording to claim 1 wherein the compound of formula (II) is of theformula (VIb):

or salts or tautomers or N-oxides or solvates thereof; wherein R²⁰ isselected from hydrogen and methyl; R^(21a) is selected from fluorine andchlorine; and R^(22a) is selected from fluorine, chlorine and methoxy.7. A combination according to claim 6 wherein the compound of formula(II) is 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof.
 8. A combination according toclaim 1 wherein the one or more other therapeutic agents are selectedfrom the following anticancer agents: topoisomerase inhibitorsalkylating agents antimetabolites DNA binders microtubule inhibitors(tubulin targeting agents) monoclonal antibodies signal transductioninhibitors radiotherapy.
 9. A combination according to claim 8 whereinthe compound of formula (II) is4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof.
 10. A combination according toclaim 8 wherein the one or more other therapeutic agents are selectedfrom the anticancer agents cisplatin, cyclophosphamide, doxorubicin,irinotecan, fludarabine, 5FU, taxanes, and mitomycin C.
 11. Acombination according to claim 10 wherein the compound of formula (II)is 4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof.
 12. A combination according toclaim 8 wherein the compound of the formula (II) and one, two, three,four or more other therapeutic agents are formulated together in adosage form containing two, three, four or more therapeutic agents. 13.A pharmaceutical composition (e.g. formulation) comprising at least onecompound of formula (II) as defined in claim 1 together with one or morepharmaceutically acceptable carriers, adjuvants, excipients, diluents,fillers, buffers, stabilisers, preservatives, lubricants, or othertherapeutic or prophylactic agents.
 14. A pharmaceutical compositioncomprising at least one compound of formula (II) as defined in claim 11together with one or more pharmaceutically acceptable carriers,adjuvants, excipients, diluents, fillers, buffers, stabilisers,preservatives, lubricants, or other therapeutic or prophylactic agents.15. A method for treating cancer, which method comprises administeringto a mammal in need thereof a combination according to claim
 8. 16. Amethod according to claim 15 wherein the compound of formula (II) andthe one or more other therapeutic agents are administered eithersimultaneously or sequentially.
 17. A method according to claim 15wherein the compound of formula (II) is4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof.
 18. A method for treating cancer,which method comprises administering to a mammal in need thereof acombination according to claim 8 wherein the compound of formula (II) is4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof, and the one or more othertherapeutic agents are selected from the anticancer agents cisplatin,cyclophosphamide, doxorubicin, irinotecan, fludarabine, 5FU, taxanes,and mitomycin C.
 19. A method for treating a disease or condition, whichmethod comprises administering to a mammal in need thereof a compound offormula (II) as defined in claim 1 for use in the treatment of a diseasewhich is: viral infections, for example herpes virus, pox virus,Epstein-Barr virus, Sindbis virus, adenovirus, HIV, HPV, HCV and HCMV;prevention of AIDS development in HIV-infected individuals; or type IIor non-insulin dependent diabetes mellitus; or autoimmune diseases; orhead trauma; or stroke; or epilepsy; or neurodegenerative disorders, forexample Alzheimer's disease, AIDS-related dementia, Parkinson's disease,amyotropic lateral sclerosis, retinitis pigmentosa, spinal muscularatropy and cerebellar degeneration or such as Alzheimer's, motor neuronedisease, progressive supranuclear palsy, corticobasal degeneration andPick's disease; or chronic inflammatory diseases, for example systemiclupus erythematosus, autoimmune mediated glomerulonephritis, rheumatoidarthritis, psoriasis, inflammatory bowel disease, and autoimmunediabetes mellitus; or cardiovascular diseases for example cardiachypertrophy, restenosis, and atherosclerosis, or such as arrhythmia; orglomerulonephritis; or myelodysplastic syndrome; or ischemic injuryassociated myocardial infarctions, stroke and reperfusion injury; ortoxin-induced or alcohol related liver diseases; or haematologicaldiseases, for example, chronic anemia and aplastic anemia; ordegenerative diseases of the musculoskeletal system, for example,osteoporosis and arthritis, or aspirin-sensitive rhinosinusitis; orcystic fibrosis; or multiple sclerosis, or kidney diseases; or cancerpain; or cancer, in particular RB+ve tumours.
 20. A method for treatinga disease or condition as defined in claim 19, which method comprisesadministering to a mammal in need thereof4-(2,6-dichloro-benzoylamino)-1H-pyrazole-3-carboxylic acidpiperidin-4-ylamide or a salt thereof.